Pharmaceutical compositions for peptide delivery

Abstract

The present invention provides a pharmaceutical composition for peptide delivery. [Solution] The present invention relates to a pharmaceutical composition comprising a pharmaceutically effective dose of at least one type of peptide in combination with a pharmaceutically acceptable dose of (a) at least one type of metal in the form of a salt or a complex, or a combination thereof, and (b) at least one type of reducing agent, wherein the at least one type of metal is selected from vanadium, chromium, and manganese, or a combination thereof, and the combination of (a) at least one type of metal in the form of a salt or a complex, or a combination thereof, and (b) at least one type of reducing agent can at least partially protect the at least one type of peptide from proteolytic degradation upon ingestion. [Selected Figure] Figure 1

Description

[Technical Field]

[0001] The present disclosure relates generally to the pharmaceutical field, and more particularly, the present invention is a pharmaceutical composition comprising a combination of a peptide with a metal salt / complex and a reducing agent, which can, at least in part, protect the peptide from proteolytic degradation upon ingestion. [Background technology]

[0002] The background material in this specification includes information that is helpful for understanding the present invention. However, the background material in this specification does not assert that the present invention is well-known art or is not relevant to the scope of protection of the present invention. Furthermore, any material or document disclosed in the background material in this specification does not expressly or implicitly represent that the present invention is well-known art.

[0003] Proteins and peptides have long been used as therapeutic and diagnostic agents, and related technologies have continued to develop rapidly to this day. However, the potential of proteins and peptides has not yet been fully understood, and their applications remain limited to parenteral injection.

[0004] Oral administration is the best and simplest route of administration. However, peptides are degraded after digestion in the gastrointestinal tract, making complete absorption difficult. Therefore, the low oral bioavailability of proteins and peptides is mainly due to enzymatic degradation in the gastrointestinal tract and the low osmolality of epithelial cells.

[0005] Many approaches have been proposed to address the problem of poor oral bioavailability of proteins and peptides, including the use of various absorption enhancers and protease inhibitors, such as soybean trypsin inhibitor, aprotinin, Bowman-Birk trypsin inhibitor, bacitracin, camostat mesilate, and amastatin (Renukuntla J et al., Int J Pharm. 2013, 447, 75-93; US Published Patent Application US20070087957A1). However, due to their toxicity and numerous side effects, no additives compatible with protease inhibitors for peptide delivery have been commercially available.

[0006] The following are a few examples of protease inhibitors that can be used for peptide delivery: (a) Soybean (i.e., soybean trypsin inhibitor): Widely considered an allergen, its availability has been limited since 1980 due to the gradual increase in the number of people suffering from soybean allergy (Moroz LA et al., N Engl J Med. 1980, 302, 1126-8; Foucard T et al., Allergy, 1999, 54, 261-5; Ramesh S, Clin Rev Allergy Immunol. 2008, 34, 217-30). Acute allergic reactions caused by soybean include coughing, sneezing, runny nose, hives, diarrhea, facial swelling, shortness of breath, tongue swelling, difficulty swallowing, drop in blood pressure, excessive sweating, fainting, anaphylactic shock, and even death. (b) Bowman (i.e., Bowman-Birk trypsin inhibitor): A soybean derivative with high oral bioavailability, even without absorption enhancers. However, oral administration of Bowman has been reported to sometimes result in unwanted systemic protease inhibition (e.g., inhibition of systemic serine proteases such as plasmin, potentially increasing the risk of thrombosis). Bowman may also induce the formation of self-antibodies (Wan XS et al., Nutr Cancer, 2002, 43, 167-73). (c) Aprotinin: Initial use at a ratio of 1:200 is known to cause allergic reactions (Mahdy AM et al., 2004, 93, 842-58), and aprotinin has been reported to be associated with a risk of acute renal failure, myocardial infarction, heart failure, stroke, and encephalopathy in cardiac / surgical patients (Mangano DT et al., N Engl J Med, 2006, 354, 353-65).

[0007] The potential health risks associated with protease inhibitors have led to their general avoidance. In addition to these limitations, protease inhibitors face challenges such as high manufacturing costs, heterogeneity, regulatory barriers, and difficulty in selective inhibition, as well as the requirement for high doses for effective activity, all of which hinder their widespread application (Renukuntla J et al., Int J Pharm. 2013, 447). Other protease inhibitors, such as bacitracin (antibiotic activity), camostat mesylate (effective in treating pancreatitis), and the inhibitor amastatin (antibacterial activity) (Renukuntla J et al., Int J Pharm. 2013, 447, 75–93; U.S. Patent Application Publication US20070087957A1), also suffer from similar side effects.

[0008] European Patent EP 3006045B1 discloses a composition of trace elements (such as copper and zinc) and a pharmaceutically acceptable reducing agent, which can optionally be supplemented with a transmucosal absorption enhancer, and which has been shown to unexpectedly have beneficial and high oral bioavailability for various peptide or protein drugs. However, due to the relationship between copper or zinc and mammalian metabolic pathways, the use of this composition in long-term treatment may result in negative interactions.

[0009] Thus, there is a need in the art for research and development of simple, safe, effective, and cost-effective pharmaceutical compositions that can deliver peptides while protecting them, at least in part, from proteolytic degradation upon ingestion. The present disclosure meets current and other needs and reduces the technical shortcomings of conventional pharmaceutical compositions and their delivery. Summary of the Invention [Problem to be solved by the invention]

[0010] The present disclosure aims to provide a pharmaceutical composition that can overcome the drawbacks associated with the compositions reported in the background art.

[0011] Another object of the present disclosure is to provide a pharmaceutical composition that effectively delivers peptides.

[0012] Another object of the present disclosure is to provide pharmaceutical compositions for oral peptide delivery.

[0013] Another object of the present disclosure is to provide pharmaceutical compositions that, at least in part, protect ingested peptides from proteolytic degradation.

[0014] Another object of the present disclosure is to provide pharmaceutical compositions that increase the oral bioavailability of peptides.

[0015] Another object of the present disclosure is to provide a safe pharmaceutical composition.

[0016] Another object of the present disclosure is to provide a pharmaceutical composition that is economically efficient to manufacture.

[0017] Another object of the present disclosure is to provide pharmaceutical compositions that are easy to prepare.

[0018] Another object of the present disclosure is to provide a pharmaceutical composition with a long shelf life.

[0019] The present disclosure relates generally to the pharmaceutical field, and specifically, the present invention relates to pharmaceutical compositions comprising a combination of a peptide with a metal salt / complex and a reducing agent, which, at least in part, protects the peptide from proteolytic degradation upon ingestion.

[0020] As an aspect of this disclosure, a pharmaceutically effective dose of at least one peptide, and a pharmaceutically acceptable dosage of (a) at least one metal in either a salt form, a complex form, or a combination thereof, in combination with (b) at least one reducing agent; The at least one metal is selected from any one or a combination of vanadium, chromium, and manganese, and the combination of (a) the at least one metal in any one or a combination of a salt and a complex, and (b) at least one reducing agent can at least partially protect the at least one peptide from proteolytic degradation upon ingestion.

[0021] In one embodiment, the at least one metal is vanadium, and the pharmaceutical composition contains a vanadium salt, a vanadium complex, or a combination thereof in an amount ranging from about 0.01 mg to about 15 mg per unit dose. In one embodiment, the vanadium salt and the vanadium complex are independently selected from the group consisting of vanadium(V) oxide, sodium vanadate, vanadium sulfate, vanadyl sulfate, vanadium biguanide, bis(maltolato)oxovanadium(IV), vanadium acetate, vanadyl picolinate, and vanadyl citrate. In one embodiment, the at least one metal is chromium, and the pharmaceutical composition contains a chromium salt, a chromium complex, or a combination thereof in an amount ranging from about 0.02 mg to about 0.5 mg per unit dose. In one embodiment, the chromium salt and the chromium complex are independently selected from the group consisting of chromium picolinate, chromium polynicotinate, chromium nicotinate, chromium chloride, and chromium acetate. In one embodiment, the at least one metal is manganese, and the pharmaceutical composition contains a manganese salt, a manganese complex, or a combination thereof, in an amount ranging from about 0.1 mg to about 10 mg per unit dose. In one embodiment, the manganese salt, or a manganese complex is independently selected from the group consisting of manganese gluconate, manganese sulfate, potassium permanganate, and manganese chloride.

[0022] In one embodiment, the at least one peptide has a molecular weight of 60 kDa or less. In one embodiment, the at least one peptide is insulin, an insulin analog, insulin lispro, insulin peglispro, insulin aspart, insulin glulisine, insulin glargine, insulin detemir, protamine-containing intermediate-acting (NPH) insulin, insulin degludec, B29K(N(ε)hexadecandioyl-γ-L-Glu) A14E B25H desB30 human insulin, B29K(N(ε)octadecandioyl-γ-L-Glu-OEG-OEG) desB30 human insulin, B29K(N(ε)octadecandioyl-γ-L-Glu) A14E B25H desB30 human insulin, B29K(N(ε)eicosandioyl-γ-L-Glu) A14E B25H desB30 human insulin, B29K(N(ε)octadecandioyl-γ-L-Glu-OEG-OEG) A14E B25H desB30 Human insulin, B29K(N(ε)eicosandioyl-γ-L-Glu-OEG-OEG) A14E B25H desB30 Human insulin, B29K(N(ε)eicosandioyl-γ-L-Glu-OEG-OEG) A14E B16H B25H desB30 Human insulin, B29K(N(ε)hexadecandioyl-γ-L-Glu) A14E B16H B25H desB30 Human insulin, B29K(N(ε)eicosandioyl-γ-L-Glu-OEG-OEG) A14E B16H B25H desB30 Human insulin, B29K(N(ε)octadecandioyl) A14E B25H desB30 Human insulin, GLP-1, GLP-1 analogues, acylated GLP-1 analogues, diacylated GLP-1 analogues, semaglutide, liraglutide, exenatide, lixisenatide,GLP-1 receptor and glucagon receptor dual agonists, amylin, amylin analogs, pramlintide, somatostatin analogs, octreotide, lanreotide, pasireotide, goserelin, buserelin, leptin, leptin analogs, metreleptin, peptide YY, peptide YY analogs, glatiramer, leuprorelin, teriparatide, desmopressin, human growth hormone, human growth hormone analogs, glycopeptides Antibiotics, glycosylated cyclic or polycyclic nonribosomal peptide antibiotics, vancomycin, teicoplanin, telavancin, bleomycin, ramoplanin, decaplanin, bortezomib, cosyntropin, chorionic gonadotropin, menotropin, sermorelin, luteinizing hormone-releasing hormone, somatropin, calcitonin, salmon calcitonin, pentagastrin, oxytocin, nesiritide, anakinra, enfuvirtide, pegviso manto, dornase alfa, lepirudin, anidulafungin, eptifibatide, interferon alfacon-1, interferon alfa-2a, interferon alfa-2b, interferon beta-1a, interferon beta-1b, interferon gamma-1b, pegylated interferon alfa-2a, pegylated interferon alfa-2b, pegylated interferon beta-1a, fibrinolysin, vasopressin, aldesleukin, epoetin alfa, darbepoetin alfa, epoetin beta, epoetin delta, epoetin omega, epoetin zeta, filgrastim, interleukin-11, cyclosporine, glucagon, urokinase, viomycin, thyrotropin-releasing hormone, leucine enkephalin, methionine enkephalin, substance P, adrenocorticotropic hormone, parathyroid hormone, or a pharmaceutically acceptable salt thereof.

[0023] In one embodiment, the at least one peptide and the at least one metal in the form of a salt, a complex, or a combination thereof are physically separated in the pharmaceutical composition. In one embodiment, the at least one peptide and the at least one metal in the form of a salt, a complex, or a combination thereof are present in separate compartments. In one embodiment, the pharmaceutical composition is present in a capsule-in-capsule dosage form or a tablet-in-capsule dosage form.

[0024] In one embodiment, the at least one reducing agent is selected from ascorbic acid, reduced glutathione, cysteine, uric acid, reducing sugars, glyceraldehyde, α-tocopherol, vitamin A, α-lipoic acid, dihydro-α-lipoic acid, glucose, galactose, lactose, maltose, thiol-containing compounds, thiomers, and pharmaceutically acceptable salts thereof, or combinations thereof. In one embodiment, the pharmaceutical composition comprises the at least one reducing agent in an amount ranging from about 1 mg to about 1000 mg per unit dose.

[0025] In one embodiment, the pharmaceutical composition further comprises at least one absorption enhancer, wherein the absorption enhancer is present in an amount ranging from about 10 mg to about 1000 mg per unit dose. In one embodiment, the pharmaceutical composition is prepared in either an oral solid or oral liquid form, and when the pharmaceutical composition is prepared in an oral liquid form, the pharmaceutical composition contains less than about 5% (v / v) water.

[0026] The contents of the present disclosure will be described in detail below based on preferred embodiments and drawings of the present disclosure, and various components, technical features, aspects and advantages of the present disclosure will be clearly disclosed. Components with the same component symbols in the drawings are the same components. [Brief explanation of the drawings]

[0027] [Figure 1]1 is a graph showing a comparison of insulin glargine (mU / L) concentration versus time from different formulations according to an embodiment of the disclosure. DETAILED DESCRIPTION OF THE INVENTION

[0028] To explain the present disclosure in detail, specific embodiments of the present disclosure will be described in detail and clearly in conjunction with the figures. The embodiments are described in detail to clearly convey the disclosure. However, the amount of detail provided is not intended to limit the anticipated variations of the embodiments. On the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the present disclosure as defined by the appended claims.

[0029] Each of the following claims defines a respective invention, and infringement claims are recognized to include equivalents to the various elements or limitations specified in the claims. Depending on the context of the following specification, the term "the present invention" as used herein may in some cases refer only to particular embodiments of the present disclosure, and in other cases, the term "the present invention" may refer to one or more, but not necessarily all, of the subject matter recited in the claims.

[0030] As used herein and in the claims, the words "a," "an," and "the" refer to plural inclusion unless the context clearly dictates otherwise. Similarly, as used herein, the term "in" includes the meanings "in" and "on," unless the context clearly dictates otherwise.

[0031] Unless otherwise stated herein or the context clearly contradicts, the methods described in this disclosure may be performed in any suitable order. The use of exemplary or exemplary terms, i.e., the term "for example," in any or all of the examples provided herein merely describes a preferred embodiment of the present disclosure and does not limit the scope of protection of the present disclosure. No language in this specification should be construed as requiring any element not recited in the claims to be essential to the practice of the present disclosure.

[0032] Various terms used below are identified below: Unless a term used in the claims is defined below, it is to be interpreted in the broadest possible light that a person of ordinary skill in the relevant art would interpret that term as reflected in the printed publications and issued patents at the time of filing.

[0033] The present disclosure relates generally to the pharmaceutical field, and specifically, the present invention is a pharmaceutical composition comprising a combination of a peptide, a metal salt / complex, and a reducing agent, which at least partially protects the peptide from proteolytic degradation upon ingestion.

[0034] Serine proteases are ubiquitous in eukaryotes and cleave peptide bonds, with the primary catalytic triad consisting of serine, histidine, and aspartic acid. Serine proteases identified in the present disclosure include trypsin, chymotrypsin, carboxypeptidase B, and aminopeptidase M. Serine proteases play a role in physiological functions in the body, particularly in digestion (proteolysis), i.e., the hydrolysis of peptide bonds and amino acids. The present disclosure aims to provide a pharmaceutical composition that includes a combination of a peptide with a metal salt / complex and a reducing agent, thereby at least partially protecting the peptide from proteolytic degradation upon ingestion.

[0035] Based on this, one aspect of the present disclosure provides a pharmaceutical composition comprising a pharmaceutically effective dose of at least one peptide in combination with a pharmaceutically acceptable dose of (a) at least one metal in the form of a salt or a complex, or a combination thereof, and (b) at least one reducing agent, wherein the at least one metal is selected from vanadium, chromium, and manganese, or a combination thereof, and the combination of (a) the at least one metal in the form of a salt or a complex, or a combination thereof, and (b) the at least one reducing agent can at least partially protect the at least one peptide from proteolytic degradation upon ingestion.

[0036] In one embodiment, the at least one metal is vanadium, and the pharmaceutical composition contains a vanadium salt, a vanadium complex, or a combination thereof in an amount ranging from about 0.01 mg to about 15 mg per unit dose. In one embodiment, the vanadium salt and the vanadium complex are independently selected from the group consisting of vanadium(V) oxide, sodium vanadate, vanadium sulfate, vanadyl sulfate, vanadium biguanide, bis(maltolato)oxovanadium(IV), vanadium acetate, vanadyl picolinate, and vanadyl citrate. In one embodiment, the at least one metal is chromium, and the pharmaceutical composition contains a chromium salt, a chromium complex, or a combination thereof in an amount ranging from about 0.02 mg to about 0.5 mg per unit dose. In one embodiment, the chromium salt and the chromium complex are independently selected from the group consisting of chromium picolinate, chromium polynicotinate, chromium nicotinate, chromium chloride, and chromium acetate. In one embodiment, the at least one metal is manganese, and the pharmaceutical composition contains a manganese salt, a manganese complex, or a combination thereof, in an amount ranging from about 0.1 mg to about 10 mg per unit dose. In one embodiment, the manganese salt, or a manganese complex is independently selected from the group consisting of manganese gluconate, manganese sulfate, potassium permanganate, and manganese chloride.

[0037] In one embodiment, the at least one peptide has a molecular weight of 60 kDa or less. In one embodiment, the at least one peptide is insulin, an insulin analog, insulin lispro, insulin peglispro, insulin aspart, insulin glulisine, insulin glargine, insulin detemir, protamine-containing intermediate-acting (NPH) insulin, insulin degludec, B29K(N(ε)hexadecandioyl-γ-L-Glu) A14E B25H desB30 human insulin, B29K(N(ε)octadecandioyl-γ-L-Glu-OEG-OEG) desB30 human insulin, B29K(N(ε)octadecandioyl-γ-L-Glu) A14E B25H desB30 human insulin, B29K(N(ε)eicosandioyl-γ-L-Glu) A14E B25H desB30 human insulin, B29K(N(ε)octadecandioyl-γ-L-Glu-OEG-OEG) A14E B25H desB30 Human insulin, B29K(N(ε)eicosandioyl-γ-L-Glu-OEG-OEG) A14E B25H desB30 Human insulin, B29K(N(ε)eicosandioyl-γ-L-Glu-OEG-OEG) A14E B16H B25H desB30 Human insulin, B29K(N(ε)hexadecandioyl-γ-L-Glu) A14E B16H B25H desB30 Human insulin, B29K(N(ε)eicosandioyl-γ-L-Glu-OEG-OEG) A14E B16H B25H desB30 Human insulin, B29K(N(ε)octadecandioyl) A14E B25H desB30 Human insulin, GLP-1, GLP-1 analogues, acylated GLP-1 analogues, diacylated GLP-1 analogues, semaglutide, liraglutide, exenatide, lixisenatide,GLP-1 receptor and glucagon receptor dual agonists, amylin, amylin analogs, pramlintide, somatostatin analogs, octreotide, lanreotide, pasireotide, goserelin, buserelin, leptin, leptin analogs, metreleptin, peptide YY, peptide YY analogs, glatiramer, leuprorelin, teriparatide, desmopressin, human growth hormone, human growth hormone analogs, glycopeptides Antibiotics, glycosylated cyclic or polycyclic nonribosomal peptide antibiotics, vancomycin, teicoplanin, telavancin, bleomycin, ramoplanin, decaplanin, bortezomib, cosyntropin, chorionic gonadotropin, menotropin, sermorelin, luteinizing hormone-releasing hormone, somatropin, calcitonin, salmon calcitonin, pentagastrin, oxytocin, nesiritide, anakinra, enfuvirtide, pegviso manto, dornase alfa, lepirudin, anidulafungin, eptifibatide, interferon alfacon-1, interferon alfa-2a, interferon alfa-2b, interferon beta-1a, interferon beta-1b, interferon gamma-1b, pegylated interferon alfa-2a, pegylated interferon alfa-2b, pegylated interferon beta-1a, fibrinolysin, vasopressin, aldesleukin, epoetin alfa, darbepoetin alfa, epoetin beta, epoetin delta, epoetin omega, epoetin zeta, filgrastim, interleukin-11, cyclosporine, glucagon, urokinase, viomycin, thyrotropin-releasing hormone, leucine enkephalin, methionine enkephalin, substance P, adrenocorticotropic hormone, parathyroid hormone, or a pharmaceutically acceptable salt thereof.

[0038] In one embodiment, the at least one peptide and the at least one metal in the form of a salt, a complex, or a combination thereof are physically separated in the pharmaceutical composition. In one embodiment, the at least one peptide and the at least one metal in the form of a salt, a complex, or a combination thereof are present in separate compartments. In one embodiment, the pharmaceutical composition is present in a capsule-in-capsule dosage form or a tablet-in-capsule dosage form.

[0039] In one embodiment, the at least one reducing agent is selected from ascorbic acid, reduced glutathione, cysteine, uric acid, reducing sugars, glyceraldehyde, α-tocopherol, vitamin A, α-lipoic acid, dihydro-α-lipoic acid, glucose, galactose, lactose, maltose, thiol-containing compounds, thiomers, and pharmaceutically acceptable salts thereof, or combinations thereof. In one embodiment, the pharmaceutical composition comprises the at least one reducing agent in an amount ranging from about 1 mg to about 1000 mg per unit dose.

[0040] In one embodiment, the pharmaceutical composition further comprises at least one absorption enhancer, wherein the absorption enhancer is present in an amount ranging from about 10 mg to about 1000 mg per unit dose. In one embodiment, the pharmaceutical composition is prepared in either an oral solid or oral liquid form, and when the pharmaceutical composition is prepared in an oral liquid form, the pharmaceutical composition contains less than about 5% (v / v) water.

[0041] In one embodiment, the peptide is any peptide or protein suitable for use as a therapeutic or diagnostic agent. In one embodiment, the peptide is a linear or cyclic peptide. In one embodiment, the peptide is a modified or derivatized peptide, such as a PEGylated peptide, a fatty acid acylated peptide, or a di-fatty acid acylated peptide. The peptide may be free of histidine residues and / or free of cysteine ​​residues. Generally, the peptide is preferably water-soluble and has at least one serine protease cleavage site, particularly at neutral pH (i.e., about pH 7), i.e., the peptide contains one or more amino acid residues suitable for or susceptible to cleavage by a serine protease (particularly a gut-derived serine protease, such as trypsin, chymotrypsin, aminopeptidase, carboxypeptidase, elastase, and / or dipeptidyl peptidase 4).

[0042] In one embodiment, the peptide is selected from the group consisting of insulin (preferably human insulin), insulin derivatives such as, but not limited to, long-acting basal insulin analogues, stabilized protease long-acting basal insulin analogues, insulin lispro, insulin peglispro, A14E B25H B29K (N(eps)octadecandioyl-gGlu-OEG-OEG), desB30 human insulin, insulin aspart, insulin glulisine, insulin glargine, insulin detemir, protamine-containing intermediate-acting (NPH) insulin, insulin degludec, and insulin derivatives such as the insulin analogues / derivatives disclosed in US Published Patent Application US20140056953A1, GLP-1, GLP-1 analogues (acylated GLP-1 analogues or diacylated GLP-1 analogues), semaglutide, liraglutide, exenatide, lixisenatide,Dual agonists of GLP-1 receptor and glucagon receptor, amylin, amylin analogs, pramlintide, somatostatin analogs (octreotide, lanreotide, or pasireotide), goserelin, buserelin, leptin, leptin analogs (metreleptin), peptide YY (PYY), PYY analogs, glatiramer (glatiramer acetate), leuprorelin, teriparatide, abaloparatide, tetracosactide, corticorelin, etelcalcetide, elcatonin, desmopressin, human growth hormone (hGH), human growth hormone analogs, glycopeptide antibiotics (vancomycin, teicoplanin, telavancin, bleomycin, ramoplanin, decaplanin, etc.), glycosylated cyclic or polycyclic nonribosomal peptide antibiotics), bortezomib, cosyntropin, chorionic gonadotropin, menotropins, sermorelin, luteinizing hormone-releasing hormone (LHRH, also known as gonadotropin-releasing hormone), somatropin, calcitonin (salmon calcitonin), pentagastrin, oxytocin, nesiritide, anakinra, enfuvirtide, pegvisomant, dornase alfa, lepirudin, anidulafungin, eptifibatide, interferon alfacon-1, interferon alpha-2a, interferon alpha-2b, interferon beta-1a, interferon beta-1b, interferon gamma-1b, pegylated interferon alpha-2a (pegylated interferon alfa-2a, pegylated interferon alfa-2b, pegylated interferon beta-1abeta-1a), fibrinolysin, vasopressin, aldesleukin, epoetin alfa, darbepoetin alfa, epoetin beta, epoetin delta, epoetin omega, epoetin zeta, filgrastim, interleukin-11, cyclosporine, glucagon, urokinase, viomycin, thyrotropin-releasing hormone (TRH), leucine enkephalin, methionine enkephalin, substance P (CAS No. 33507-63-0), adrenocorticotropic hormone (ACTH), insulin analogs such as parathyroid hormone (PTH), or pharmaceutically acceptable salts thereof, or combinations thereof. However, any other peptide molecule known or recognized by those skilled in the art can be utilized to fulfill its intended purpose without departing from the scope and spirit of the present invention as presented in this disclosure.

[0043] In one embodiment, the human subject peptide is selected from any of endogenous peptides, such as insulin or glucagon, or a combination thereof. In a preferred embodiment, the corresponding human isoform of the peptide is used, obtained by recombinant expression or chemical synthesis. However, any other human isoform peptide known or recognized by those skilled in the art may be utilized to fulfill its intended purpose without departing from the scope and spirit of the present invention as presented in this disclosure.

[0044] In one embodiment, the peptide is an insulin analog. In one embodiment, the insulin analog is selected from insulin detemir, insulin glargine, insulin degludec, and other insulin analogs of human, porcine, or fish origin, or a combination thereof. However, any other insulin analog / derivative known or recognized by those skilled in the art may be utilized to serve its intended purpose without departing from the scope and spirit of the present invention as presented in this disclosure.

[0045] In one embodiment, a mixture of two or more peptides can be used. In one embodiment, a mixture of human insulin and a GLP-1 agonist (e.g., liraglutide, semaglutide, exenatide, or lixisenatide) can be used. However, any mixture of two or more peptides (including the peptides described above) known or recognized by those skilled in the art can be used to achieve its intended purpose without departing from the scope and spirit of the present invention as presented in this disclosure.

[0046] In one embodiment, at least one peptide has a molecular weight of 60 kDa or less. In one embodiment, at least one peptide has a molecular weight of 40 kDa or less. In one embodiment, at least one peptide has a molecular weight of 30 kDa or less. In one embodiment, at least one peptide has a molecular weight of 20 kDa or less. In one embodiment, at least one peptide has a molecular weight of 10 kDa or less. In one embodiment, at least one peptide has a molecular weight of 300 Da or more to 50 kDa or less. However, peptides having any range of molecular weight known or recognized by those of skill in the art can be utilized to fulfill their intended purposes without departing from the scope and spirit of the present invention as presented in this disclosure.

[0047] In one embodiment, the molecular weight of at least one peptide can be measured by any method known or recognized by one of skill in the art to serve its intended purpose without departing from the scope and spirit of the present invention as presented in this disclosure, such as mass spectrometry (e.g., electrospray ionization mass spectrometry (ESI-MS) or matrix-assisted laser desorption / ionization mass spectrometry (MALDI-MA)), gel electrophoresis (e.g., sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)), hydrodynamic methods (e.g., gel filtration chromatography or gradient sedimentation), or static light scattering (e.g., multi-angle light scattering detector (MALS)), etc.

[0048] In one embodiment, the at least one metal is vanadium, and the pharmaceutical composition comprises a vanadium salt and / or complex, independently selected from the group consisting of vanadium(IV), vanadium(V) such as vanadyl (VO2) salt complexes, and vanadium as the anion in a vanadate / complex. In one embodiment, the vanadium salts and vanadium complexes are selected from any one or combination of vanadium(V) oxide, vanadium pentoxide, vanadium dioxide, sodium vanadate, vanadium sulfate, vanadyl sulfate, sodium metavanadate, vanadium tetrachloride, vanadium(V) oxychloride, vanadium oxytrichloride, vanadyl chloride, vanadium trichloride oxide, ammonium vanadate, ammonium vanadium oxide, vanadium monosulfide, vanadium sulfide, vanadium(IV) chloride, vanadium biguanide, bis(maltolato)oxovanadium(IV), vanadium acetate, vanadyl picolinate, vanadyl citrate, and the like. In one embodiment, the vanadium salts and vanadium complexes are vanadium(V). The use of vanadium(V) salts and complexes is advantageous compared to vanadium(IV) salts and complexes due to their good water solubility and better oxidation state stability. In one embodiment, the vanadium salts and complexes are vanadium (IV) salts and / or complexes in which vanadium is part of the anion, as vanadate, or part of the cation, as vanadyl, although any vanadium salts and complexes or combinations thereof known or recognized by those of ordinary skill in the art may be utilized to serve their intended purposes without departing from the scope and spirit of the present invention as presented in this disclosure.

[0049] In one embodiment, the pharmaceutical composition contains either a vanadium salt or a vanadium complex, or a combination thereof, in an amount ranging from about 0.01 mg to about 15 mg per unit dose.

[0050] In one embodiment, the chromium salts and chromium complexes are preferably selected from chromium(III) salts and / or complexes. In one embodiment, any one or combination of chromium salts and chromium complexes is selected from chromium picolinate, chromium chloride, chromium nicotinate, chromium polynicotinate, chromium acetate, trivalent chromium, high chromium yeast, chromium pyridine-2-carboxylate, chromium tripicolinate, chromium 2-pyridinecarboxylate, tris(picolinato)chromium, and the like. In one embodiment, the chromium salts and chromium complexes are more preferably selected from chromium picolinate, chromium polynicotinate, chromium nicotinate, chromium chloride, chromium acetate, and the like. However, any chromium salts and complexes or combinations thereof known or recognized by those skilled in the art can be utilized to fulfill their intended purposes without departing from the scope and spirit of the present invention as presented in this disclosure.

[0051] In one embodiment, the pharmaceutical composition contains either a chromium salt or a chromium complex, or a combination thereof, in an amount ranging from about 0.02 mg to about 0.5 mg per unit dose.

[0052] In one embodiment, the manganese salts and / or complexes, or combinations thereof, are selected from manganese(II) salts and / or complexes, manganese(III) salts and / or complexes, and manganese salts and / or complexes such as permanganate (VO). In one embodiment, the manganese salts and / or complexes, or combinations thereof, are selected from manganese(II) sulfate (MnSO), manganese(II) chloride (MnCl), manganese(III) acetate, potassium permanganate, sodium permanganate, manganese gluconate, and the like. In one embodiment, the manganese salts and / or complexes are more preferably selected from manganese(III) salts and / or complexes. In one embodiment, the manganese(III) salts and / or complexes are manganese gluconate, manganese sulfate, manganese chloride, and the like, or combinations thereof. However, any chromium salts and complexes or combinations thereof known or recognized by those skilled in the art may be utilized to serve its intended purpose without departing from the scope and spirit of the present invention as presented in this disclosure.

[0053] In one embodiment, the pharmaceutical composition contains either a manganese salt or a manganese complex, or a combination thereof, in an amount ranging from about 0.01 mg to about 50 mg, preferably from about 0.1 mg to about 10 mg, per unit dose.

[0054] In one embodiment, any vanadium, chromium, manganese salts and complexes known or recognized by those skilled in the art may be utilized to serve their intended purposes without departing from the scope and spirit of the present invention as presented in this disclosure.

[0055] In one embodiment, vanadium salts and complexes are preferred over chromium and manganese salts and complexes because vanadium salts and complexes can significantly increase the oral bioavailability of peptides. In one embodiment, chromium salts and complexes are preferred over manganese salts and complexes. In one embodiment, the use of chromium salts and complexes is advantageous in that they are less toxic. In one embodiment, the use of manganese salts and complexes is superior to vanadium and chromium salts and complexes in that manganese salts and complexes are safe for humans even at high doses.

[0056] In one embodiment, the reducing agent is selected from any one or combination of ascorbic acid (preferably an ascorbate salt such as sodium ascorbate), reduced glutathione (GSH), cysteine, uric acid, reducing sugars (reducing monosaccharides such as glucose, glyceraldehyde, or galactose, or reducing disaccharides such as lactose or maltose), mannitol, α-tocopherol, vitamin A, α-lipoic acid, dihydro-α-lipoic acid (DHLA), thiol-containing compounds, thiomers (including the thiomers disclosed in Laffleur F et al., Future Med Chem, 2012, 4, 2205-16), and the like. In one embodiment, a mixture of two or more reducing agents can be used, with ascorbate and reduced glutathione being preferred. However, any reducing agent or combination thereof known or recognized by those skilled in the art can be utilized to achieve its intended purpose without departing from the scope and spirit of the present invention as presented in this disclosure.

[0057] In one embodiment, the pharmaceutical composition contains the reducing agent in an amount ranging from about 1.0 mg to about 1000 mg, preferably from about 50 mg to about 500 mg, per unit dose.

[0058] In one embodiment, the pharmaceutical composition further comprises at least one absorption enhancer (or permeation enhancer). It should be noted that, in this disclosure, the terms "absorption enhancer" and "permeation enhancer," which are used interchangeably and synonymously, include absorption enhancers and permeation enhancers known or recognized by those skilled in the art. In one embodiment, administration of at least one absorption enhancer or permeation enhancer improves or enhances the mucosal absorption rate of the peptide in the gastrointestinal tract, particularly when the peptide is large in size. In one embodiment, the at least one absorption enhancer or permeation enhancer is selected from either a zwitterionic absorption enhancer or a non-ionic absorption enhancer, or a combination thereof. In one embodiment, the at least one enhancer is a C8-C20 alkanoylcarnitine (preferably lauroylcarnitine, myristoylcarnitine, or palmitoylcarnitine, such as lauroylcarnitine chloride, myristoylcarnitine chloride, or palmitoylcarnitine chloride), salicylic acid (preferably a salicylate, such as sodium salicylate), a salicylic acid derivative (e.g., 3-methoxysalicylic acid, 5-methoxysalicylic acid, or homovanillic acid), a C8-C20 alkanoic acid (preferably a C8-C20 alkanoate, more preferably a caprate, caprylate, myristate, palmitate, such as sodium caprate, sodium caprylate, sodium myristate, sodium palmitate, or sodium stearate), or a C8-C20 alkanoic acid (preferably a C8-C20 alkanoate, more preferably a caprate, caprylate, myristate, palmitate, or sodium stearate). salt or stearate), citric acid (preferably a citrate such as sodium citrate), fatty acid acylated amino acids (fatty acid acylated amino acids disclosed in U.S. Patent Application Publication US20140056953A1, sodium lauroyl alaninate, N-dodecanoyl-L-alanine, sodium lauroyl asparaginate, N-dodecanoyl-L-asparagine, lauroyl aspartic acid, N-dodecanoyl-L-aspartic acid, sodium lauroyl cysteinate, N-dodecanoyl L-cysteine, sodium lauroyl glutamic acid, N-dodecanoyl-L-glutamic acid, sodium lauroyl glutamate, N-dodecanoyl L-glutamine, sodium lauroyl glycinate, N-dodecanoyl-L-glycine, sodium lauroyl histidinate,N-Dodecanoyl-L-histidine, Sodium Lauroyl Isoleucinate, N-Dodecanoyl L-Isoleucine, Sodium Lauroyl Leucinate, N-Dodecanoyl L-Leucine, Sodium Lauroyl Methioninate, N-Dodecanoyl L-Methionine, Sodium Lauroyl Phenylalaninate, N-Dodecanoyl-L-Phenylalanine, Sodium Lauroyl Prolinate, N-Dodecanoyl L-Proline, Sodium Lauroyl Serinate, N-Dodecanoyl L-Serine, Sodium Lauroyl Threoninate, N-Dodecanoyl N-L-Threonine, Sodium Lauroyl Tryptophanate, N-Dodecanoyl-L-Tryptophan, Sodium Lauroyl Tyrosinate, N-Dodecanoyl-L-Tyrosine, Sodium Lauroyl Valinate, N-Dodecanoyl-L-Valine, Sodium Lauroyl Sarcosinate, N-Dodecanoyl-L-Sarcosine, Sodium Caprylic Alaninate, N-Decanoyl-L-Alanine, Sodium Caprylic Asparaginate, N-Decanoyl-L-Asparagine, Sodium Caprylic Aspartate, N-Decanoyl-L-Asparagine Acid, Sodium Capric Cysteinate, N-Decanoyl-L-Cysteine, Sodium Capric Glutamate, N-Decanoyl-L-Glutamic Acid, Sodium Capric Glutaminate, N-Decanoyl-L-Glutamine, Sodium Capric Glycinate, N-Decanoyl-L-Glycine, Sodium Capric Histidinate, N-Decanoyl-L-Histidine, Sodium Capric Isoleucinate, N-Decanoyl-L-Isoleucine, Sodium Capric Leucinate, N-Decanoyl-L-Leucine, Sodium Caprylate Capric methioninate, N-decanoyl-L-methionine, sodium capric phenylalaninate, N-decanoyl-L-phenylalanine, sodium capric prolinate, N-decanoyl-L-proline, sodium capric serinate, N-decanoyl-L-serine, sodium capric threoninate, N-decanoyl-L-threonine, sodium capric tryptophanate, N-decanoyl-L-tryptophan, sodium capric tyrosinate, N-decanoyl-L-tyrosine, sodium capric valinate,N-Decanoyl-L-valine, Sodium Capric Sarcosinate, N-Decanoyl-L-sarcosine, Sodium Oleoyl Sarcosinate, Sodium N-Decyl Leucine, Sodium Stearoyl Glutamate (Amisoft HS-11 P), Sodium Myristoyl Glutamate (Amisoft MS-11), Sodium Lauroyl Glutamate (Amisoft LS-11), Sodium Cocoyl Glutamate (Amisoft CS-11), Sodium Cocoyl Glycinate (Amilite GCS-11), Sodium N-Decyl Leucine, Sodium Cocoyl Glycine, Sodium Cocoyl Glutamate, Sodium Lauroyl Alaninate, N-Dodecanoyl L-Alanine, Sodium Lauroyl Asparaginate, N-Dodecanoyl L-Asparagine, Sodium Lauroyl Aspartate, N-Dodecanoyl L-Aspartic Acid, Sodium Lauroyl Cysteinate, N-Dodecanoyl L-Cysteine, Sodium Lauroyl Glutamate, N-Dodecanoyl L-Glutamic Acid, Sodium Lauroyl Glutaminate, N-Dodecanoyl L-Glutamine, Sodium Lauroyl Glycinate, N-Dodecanoyl L-Glycine, Sodium Lauroyl Histidinate, N-Dodecanoyl-L-Histidine, Sodium Lauroyl Isoleucinate, N-Dodecanoyl L-Isoleucine, Sodium Lauroyl Leucine N-dodecanoyl L-leucine, sodium lauroyl methioninate, N-dodecanoyl L-methionine, sodium lauroyl phenylalaninate, N-dodecanoyl-L-phenylalanine, sodium lauroyl prolinate, N-dodecanoyl L-proline, sodium lauroyl serinate, N-dodecanoyl L-serine, sodium lauroyl threoninate, N-dodecanoyl-L-threonine, sodium lauroyl tryptophanate, N-dodecanoyl-L-tryptophan, sodium lauroyl tyrosinate, N-dodecanoyl-L-tyrosine, sodium lauroyl valinate, N-dodecanoyl L-valine, N-dodecanoyl-L-sarcosine, sodium capric alaninate, N-decanoyl-L-alanine, sodium capric asparaginate, N-decanoyl-L-asparagine,Sodium capric aspartate, N-decanoyl-L-aspartic acid, sodium capric cysteinate, N-decanoyl-L-cysteine, sodium capric glutamate, N-decanoyl-L-glutamic acid, sodium capric glutamate, N-decanoyl-L-glutamine, sodium capric glycinate, N-decanoyl-L-glycine, sodium capric histidinate, N-decanoyl-L-histidine, sodium capric isoleucinate, N-decanoyl-L-isoleucine, sodium capric leucinate, N-decanoyl-L-leucine, sodium capric methioninate, N-decanoyl-L-methionine, sodium capric phenylalaninate, N-decanoyl-L-phenylalanine, sodium capric prolinate, N-decanoyl-L-proline, sodium capric serine hydroxybenzoates, N-decanoyl-L-serine, sodium capric threoninate, N-decanoyl-L-threonine, sodium capric tryptophanate, N-decanoyl-L-tryptophan, sodium capric tyrosinate, N-decanoyl-L-tyrosine, sodium capric valinate, N-decanoyl-L-valine, sodium capric sarcosinate, sodium oleoyl sarcosinate, and pharmaceutically acceptable salts of the above-mentioned compounds such as C8 to C20 alkanoyl sarcosinates (lauroyl sarcosinates such as sodium lauroyl sarcosinate), or compounds in which one amino acid out of the 20 standard α-amino acids that constitute proteins is acylated with a C8 to C20 alkanoic acid, but are not limited to these), alkyl saccharides (for example, C8 to C10 alkyl polysaccharides, specifically Multitrope, TM1620-LQ-(MV), C1-C20 alkyl saccharides such as n-octyl-β-D-glucopyranoside or n-dodecyl-β-D-maltoside), cyclodextrins (α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, methyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin or sulfobutylether-β-cyclodextrin), sodium N-[8-(2-hydroxybenzoyl)amino]caprylate (SNAC), thiomers (including the thiomers disclosed by Laffleur F et.al., Future Med Chem. 2012, 4, 2205-16), calcium chelating compounds (ethylenediaminetetraacetic acid (EDTA), glycoletherdiaminetetraacetic acid (EGTA), sodium citrate and polyacrylic acid), Cremophor EL (Kolliphor EL; CAS no.61791-12-6), chitosan, N,N,N-trimethylchitosan, benzalkonium chloride, bestatin, cetylpyridinium chloride, cetyltrimethylammonium bromide, C2-C20 alkanols (such as ethanol, decanol, lauryl alcohol, myristyl alcohol, or palmityl alcohol), C8-C20 alkenols (such as oleyl alcohol), C8-C20 alkenoic acids (such as oleic acid), dextran sulfate, diethylene glycol monoethyl ether (transcutol), 1-dodecylazacycloheptan-2-one (Azone®), ethyl caprylate, glyceryl monolaurate, lysophosphatidylcholine, menthol, C8-C20 alkylamines, C8-C20 alkenylamines (such as oleylamine), phosphatidylcholine, poloxamer, polyethylene glycol monolaurate, polyoxyethylene, polypropylene monolaurate The surfactant may be selected from any one of pyrene glycol, polysorbate (polysorbate 80), deoxycholic acid (sodium deoxycholate), sodium glycocholate, sodium glycodeoxycholate, sodium lauryl sulfate (SDS), taurocholic acid (such as sodium taurocholate), taurodeoxycholic acid (such as sodium taurodeoxycholate), sucrose laurate, sulfoxides (C1-C10 alkyl-C1-C10 alkyl-sulfoxides such as decylmethyl sulfoxide or dimethyl sulfoxide), cyclopentadecalactone, 8-(N-2-hydroxy-5-chloro-benzoyl)-amino-caprylic acid (5-CNAC), dodecyl-2-N,N-dimethylaminopropionic acid (DDAIP), D-α-tocopheryl polyethylene glycol 1000 succinate (TPGS), and pharmaceutically acceptable salts of the above compounds, or a combination thereof. In one embodiment, a mixture of any two or more absorption enhancers, including those described above, can be used, although any absorption enhancer or combination thereof known or recognized by those of ordinary skill in the art can be utilized to achieve its intended purpose without departing from the scope and spirit of the present invention as presented in this disclosure.

[0059] In one embodiment, the pharmaceutical composition optionally comprises an absorption enhancer or permeation enhancer in an amount ranging from about 10 mg to about 1000 mg, preferably from about 50 mg to about 500 mg, per unit dose.

[0060] In one embodiment, the pharmaceutical composition is formulated such that when the pharmaceutical composition is added to 10 milliliters of a 5% hydrochloric acid solution, the acid is neutralized to provide a pH greater than about 6. In one embodiment, the pharmaceutical composition is formulated such that when the pharmaceutical composition is added to 10 milliliters of an aqueous solution, the pH is in the range of 6 to 9.

[0061] In one embodiment, a pharmaceutical composition comprising a pharmaceutically effective dose of at least one peptide having a molecular weight of 50 kDa or less, a vanadium salt and / or a vanadium complex or a composition thereof, at least one reducing agent, and optionally an absorption enhancer, is orally administered and can at least partially protect the at least one peptide from proteolytic degradation upon ingestion.

[0062] In one embodiment, a pharmaceutical composition comprising a pharmaceutically effective dose of at least one peptide having a molecular weight of 50 kDa or less, a chromium salt and / or a chromium complex, at least one reducing agent, and optionally an absorption enhancer, is orally administered and can at least partially protect the at least one peptide from proteolytic degradation upon ingestion.

[0063] In one embodiment, a pharmaceutical composition comprising a pharmaceutically effective dose of at least one peptide having a molecular weight of 50 kDa or less, a manganese salt and / or a manganese complex, at least one reducing agent, and optionally an absorption enhancer, is orally administered and can at least partially protect the at least one peptide from proteolytic degradation upon ingestion.

[0064] In one embodiment, the pharmaceutical composition optionally further comprises any one or a combination of one or more pharmaceutically acceptable excipients such as carriers, diluents, fillers, disintegrants, lubricants, binders, colorants, pigments, stabilizers, preservatives, antioxidants, and / or solubility enhancers. In one embodiment, the pharmaceutical composition optionally further comprises one or more pharmaceutically acceptable additives such as vitamin E, histidine, microcrystalline cellulose (MCC), mannitol, starch, sorbitol, and / or lactose. In one embodiment, the pharmaceutical composition can be adjusted by any technique known or appreciated by those skilled in the art to achieve its intended purpose without departing from the scope and spirit of the invention as presented in this disclosure.

[0065] In one embodiment, the at least one solubility enhancer is selected from the group consisting of polyethylene glycol, ethylene glycol, propylene glycol, nonionic surfactants, tyloxapol, polysorbate 80, macrogol-15-hydroxystearate, phospholipids, lecithin, dimyristoylphosphatidylcholine, dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, cyclodextrin, α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, hydroxyethyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin, hydroxyethyl-γ-cyclodextrin, hydroxypropyl-γ-cyclodextrin, dihydroxypropyl-β-cyclodextrin, sulfobutyl ester, hydroxypropyl ... The preferred dissolution enhancers are selected from any one or combination of butyl-β-cyclodextrin, sulfobutylether-γ-cyclodextrin, glucosyl-α-cyclodextrin, glucosyl-β-cyclodextrin, diglucosyl-β-cyclodextrin, maltosyl-α-cyclodextrin, maltosyl-β-cyclodextrin, maltosyl-γ-cyclodextrin, maltotriosyl-β-cyclodextrin, maltotriosyl-γ-cyclodextrin, dimaltosyl-β-cyclodextrin, methyl-β-cyclodextrin, carboxyalkyl thioether, hydroxypropylmethylcellulose, hydroxypropylcellulose, polyvinylpyrrolidone, vinyl acetate copolymer, vinylpyrrolidone, sodium lauryl sulfate, or dioctyl sodium sulfosuccinate, etc. However, any dissolution enhancer or combination thereof as known or recognized by those skilled in the art may be utilized to achieve its intended purpose without departing from the scope and spirit of the present invention as presented in this disclosure.

[0066] In one embodiment, the pharmaceutical composition is prepared for oral administration, preferably for oral administration, in which at least one peptide, at least one metal in the form of a metal salt or a metal complex, or a combination thereof, at least one reducing agent, and optionally an absorption enhancer are administered orally.

[0067] In one embodiment, the oral pharmaceutical composition dosage form is selected from any one or combination of tablets (coated or uncoated), capsules (soft gelatin capsules, hard gelatin capsules, HPMC capsules, or HPMCP capsules), capsule-in-capsules, tablet-in-capsules, lozenges / troches, suppositories, solutions, emulsions, suspensions, syrups, elixirs, redissolvable powders and granules, dispersible powders and granules, medicated gums, chewable tablets, effervescent tablets, multiparticulate dosage forms, and the like, although any pharmaceutical composition dosage form or combination thereof known or recognized by one of ordinary skill in the art may be utilized to fulfill its intended purpose without departing from the scope and spirit of the present invention as presented in this disclosure.

[0068] In one embodiment, the tablet may contain any one or a combination of excipients such as, but not limited to, microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, calcium hydrogen phosphate, glycine, disintegrants (e.g., starch, preferably corn, potato, or tapioca starch), sodium starch glycolate, croscarmellose sodium, and certain complex silicates, as well as granulation aids (e.g., polyvinylpyrrolidone, hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), sucrose, gelatin, acacia gum, etc.), lubricants (e.g., magnesium stearate, stearic acid, glyceryl behenate, talc powder, etc.), however, any excipient or combination thereof known or recognized by one of ordinary skill in the art may be utilized to accomplish its intended purpose without departing from the scope and spirit of the present invention as presented in this disclosure.

[0069] In one embodiment, the capsule may contain any one or combination of excipients such as, but not limited to, lactose, starch, cellulose, or high molecular weight polyethylene glycols, although any excipient or combination thereof known or recognized by one of ordinary skill in the art may be utilized to serve its intended purpose without departing from the scope and spirit of the present invention as presented in this disclosure.

[0070] In one embodiment, aqueous suspensions and / or elixirs may contain any one or combination of excipients such as, but not limited to, sweeteners or flavorings, colorings or dyes, emulsifying and / or suspending agents, and diluents (such as water, ethanol, propylene glycol, and glycerin), although any excipient or combination thereof known or recognized by those skilled in the art may be utilized to accomplish its intended purpose without departing from the scope and spirit of the present invention as presented in this disclosure.

[0071] In one embodiment, the pharmaceutical composition is prepared in either an oral solid dosage form or an oral liquid dosage form, and when the pharmaceutical composition is prepared in the form of an oral liquid dosage form, the pharmaceutical composition contains less than about 5% (v / v), preferably less than 3% (v / v), more preferably less than 1% (v / v), even more preferably less than 0.5% (v / v), even more preferably less than 0.1% (v / v), and even more preferably no water. In one embodiment, oral liquid dosage forms are particularly advantageous in that they can enhance storage stability. In an alternative embodiment, oral liquid dosage forms can be prepared quickly before administration, and long-term storage should be avoided.

[0072] The amounts of vanadium, chromium, and / or manganese utilized in accordance with the embodiments of the present disclosure are far lower than the recommended daily intake of these trace elements and therefore can be considered safe. Furthermore, the combination of vanadium, chromium, and / or manganese with a reducing agent not only inhibits the action of serine proteases in the gastrointestinal tract but also has no systemic effects, resulting in further improved safety compared to the protease inhibitors described in the background art above. Furthermore, compared to the protease inhibitors proposed for oral delivery of peptide or protein drugs described in the background art above, vanadium, chromium, and / or manganese, and a reducing agent such as ascorbic acid or reduced glutathione, can be provided at significantly lower production costs.

[0073] Typically, a physician will determine the actual optimal dosage for each individual patient. The specific dosage and frequency of administration for any individual patient will vary and depend on a variety of factors, including the activity of the specific peptide or protein used, the metabolic stability and response time of the specific peptide or protein compound, as well as the individual patient's age, weight, health, sex, diet, method and time of administration, excretion rate, pharmaceutical composition, the severity of the particular condition, and the individual patient's treatment status. The exact dosage is ultimately at the discretion of the attending physician or veterinarian. The individual or patient to be treated, such as an individual or patient in need of treatment or prophylaxis, can be an animal (e.g., a non-human animal), vertebrate, mammal, rodent (e.g., a guinea pig, hamster, rat, or mouse), murine (e.g., a mouse), canine (e.g., a dog), feline (e.g., a cat), suidae (e.g., a pig), equine (e.g., a horse), primate, ape (e.g., a monkey or ape), monkey (e.g., a gorilla, chimpanzee, orangutan, gibbon), or human. The present invention also contemplates treating economically or agriculturally important animals. Non-limiting examples of agriculturally important animals are sheep, cattle, and pigs, although cats and dogs, for example, may also be considered economically important animals. The individual / patient is preferably a mammal, more preferably a human or non-human mammal (such as a guinea pig, hamster, rat, mouse, rabbit, dog, cat, horse, monkey, ape, marmoset, baboon, gorilla, chimpanzee, orangutan, gibbon, sheep, cow, or pig), and even more preferably the patient / individual is human.

[0074] In one embodiment, the at least one peptide, the at least one metal in the form of a metal salt or complex, or a combination thereof, the at least one reducing agent, and the optional absorption enhancer can be administered simultaneously, concomitantly, or sequentially. In one embodiment, sequential administration involves first administering the at least one metal in the form of a metal salt or complex, or a combination thereof, and the at least one reducing agent, followed by the at least one peptide and the optional absorption enhancer (e.g., at least about 5 minutes after the first administration, preferably about 5 minutes to about 3 hours after the first administration, and more preferably about 10 minutes to about 1 hour after the first administration), which is particularly advantageous when the peptide is insulin (human insulin). In one embodiment, at least one metal in the form of a metal salt or complex, or a combination thereof, at least one reducing agent, and an optional absorption enhancer are administered, followed by administration of the peptide (e.g., at least about 5 minutes after the first administration, preferably about 5 minutes to 3 hours after the first administration, and more preferably about 10 minutes to 1 hour after the first administration), which is also advantageous when the peptide is insulin (human insulin). In one embodiment, the at least one metal is selected from any one of vanadium, chromium, and manganese, or a combination thereof.

[0075] In one embodiment, the simultaneous administration can be in the same pharmaceutical composition, or in two or more different / separate pharmaceutical compositions, or in two or more different / separate compartments of the same pharmaceutical dosage form, with at least one metal in the form of a metal salt and / or complex, at least one reducing agent administered first, followed by at least one peptide and optional absorption enhancer. In one embodiment, the at least one metal is vanadium, chromium, and / or manganese, or a combination thereof.

[0076] In one embodiment, the at least one peptide and the at least one metal in either a salt form, a complex form, or a combination thereof are physically separated in the pharmaceutical composition.

[0077] In one embodiment, the pharmaceutical dosage form comprises at least two separate compartments that are physically separated from one another (e.g., via a physical separating layer). In one embodiment, the pharmaceutical dosage form comprises a physical separating layer between at least one peptide and at least one metal in the form of a salt, a complex, or a combination thereof. In one embodiment, in the pharmaceutical dosage form, the at least one peptide is present only in a first compartment, and the at least one metal in the form of a salt, a complex, or a combination thereof is present only in a second compartment. In one embodiment, in the pharmaceutical dosage form, the reducing agent can be present either in the first compartment, or in the second compartment, or in both the first and second compartments, or in a third compartment. In one embodiment, the at least one metal is selected from vanadium, chromium, and manganese, or a combination thereof.

[0078] In one embodiment, the pharmaceutical composition is in a capsule-in-capsule dosage form or a tablet-in-capsule dosage form, and the pharmaceutical composition comprises at least one peptide having a molecular weight of 50 kDa or less, present in a first compartment of the pharmaceutical dosage form; at least one metal in the form of a salt or complex of the metal, or a combination thereof, present in a second compartment of the dosage form; and a reducing agent present in the first and / or second compartment of the dosage form.

[0079] In one embodiment, the present invention provides a pharmaceutical dosage form (e.g., a multiparticulate pharmaceutical dosage form) comprising: at least one peptide having a molecular weight of 60 kDa or less present in a first compartment of the pharmaceutical dosage form; at least one reducing agent present in a second compartment of the dosage form; and at least one metal in the form of a metal salt or complex, or a combination thereof, present in a third compartment of the dosage form. In one embodiment, the pharmaceutical dosage form is a capsule-in-capsule dosage form or a tablet-in-capsule dosage form. In one embodiment, when the pharmaceutical dosage form is a capsule-in-capsule dosage form, the larger outer capsule (whose contents are released first) contains at least one metal in the form of a metal salt or complex, or a combination thereof, and the reducing agent, and the smaller inner capsule (whose contents are released later) contains the peptide. In one embodiment, the at least one metal is selected from vanadium, chromium, and manganese, or a combination thereof. In one embodiment, the pharmaceutical dosage form is a modified release dosage form (such as a dosage form with an enteric coating, such as a capsule, multiparticulate, or tablet), a dosage form coated with Eudragit L30D55 ​​or Eudragit FS30D, such as a capsule, multiparticulate, or tablet, an HPMCP capsule (commercially known as AR Capsules), or a modified release dosage form (such as a capsule, multiparticulate, or tablet) with Eudragit L30D55 ​​or Eudragit FS30D. (R) However, any other agent known or recognized by those skilled in the art would depart from the scope and spirit of the present invention as presented in this disclosure. [Example]

[0080] Serine proteases: trypsin, chymotrypsin, carboxypeptidase B, and aminopeptidase M. Serine proteases are responsible for the proteolytic cleavage of peptide bonds and amino acids. This experiment tested the oxidative inactivation of serine proteases under different combinations of metal ions and reducing agents.

[0081] Enzyme activity test: Using ultraviolet spectroscopy under specific wavelengths, the enzyme activity of different enzymes in specific substrates was measured. This test result was used as a negative control. The enzyme activity calculation formula is as follows:

number

[0082] Enzyme inhibition test: Incubation was carried out in the presence of inhibitors of each enzyme, and the test results were used as positive controls.

[0083] Incubation in the presence of metal ions and reducing agents: Combinations of enzyme, metal salt, and reducing agent were incubated with the enzyme for a set period of time in a 96-well microtiter plate to test the oxidative inactivation of the enzyme in the presence of substrate. Enzyme inactivation in the presence of metal ions and reducing agents was compared to the original enzyme activity in the presence of substrate as a negative control and to a positive control in the presence of inhibitor.

[0084] pH measurement: A Hanna combination pH electrode was used to observe the change in pH value after incubating the enzyme in a combination of metal ions and a reducing agent.

[0085] Zymography: Zymography was performed on the tested enzymes. Zymography is an electrophoretic technique that detects hydrolytic enzymes based on their substrate repertoire. In other words, when an acrylamide gel is prepared, the enzyme's substrate is incorporated into the gel layer to be analyzed, and the digestion of the substrate by the enzyme is observed.

[0086] Kit analysis: Enzyme inactivation was verified using a protease fluorescence assay kit and a trypsin activity assay kit.

[0087] material:

[0088] Table 1A below lists test materials for evaluating the inhibitory effect of different metals and metal complexes (optionally in combination with one or more reducing agents) on serine protease activity. [Table 1A] TIFF0007744741000003.tif154156

[0089] Table 1B below shows the combinations of different metal salts / metal complexes and reducing agents, and the inhibitory effects on serine proteases were evaluated. [Table 1B]

[0090] Measurement of trypsin enzyme activity using Nα-benzoyl-L-arginine ethyl ester (BAEE)

[0091] A 200 unit / mL cold HCl solution of trypsin and a 0.25 mM BAEE substrate solution were prepared and incubated. The above prepared solutions were inverted and mixed to form reaction mixtures. A of the blank solution (without enzyme) and the test solution (reaction mixture) were 253The increase in absorbance at 1000 kJ / min was measured and recorded (using a minimum of four data points over one minute). The maximum linear rate was used to measure the increase in absorbance at 1000 kJ / min for both the blank and test solutions. 253 / min. The formula for the 3 ml trypsin test is as follows:

number

[0092] Measurement of trypsin inactivation in the presence of inhibitors: Trypsin was incubated at 1 mM / L with specific (known) inhibitors (listed in Table 1A) and metal salt / reducing agent combinations (listed in Table 2 below). Inactivation by inhibitors served as a positive control. [Table 2]

[0093] Determination of oxidative inactivation (activity and pH) of trypsin in the presence of combinations of metal ions and reducing agents.

[0094] For a 200 μl reaction mixture, approximately 10 μl of trypsin was incubated in a 96-well microtiter plate at 37°C with a combination of metal salts and reducing agents (listed in Table 2, serial numbers 1 to 31) at the respective concentrations in buffer (Table 3 below provides details regarding the use of specific combinations from those provided in Table 2, serial numbers 1 to 31). This was followed by the addition of 90 μl of substrate (listed in Table 1A). Enzyme activity was measured spectrophotometrically with a microplate reader and compared to the original activity assay. pH was measured with a Hanna Combination pH electrode (reactions with the enzyme were performed in triplicate). Table 4 below shows the enzymatic activity of trypsin after specific time periods following treatment with the metal salt / complex and reducing agent combinations under evaluation. [Table 3] [Table 4]

[0095] Tables 5A to 5C below show the pH values ​​measured for trypsin at different time intervals (i.e., 5 minutes, 15 minutes, and 30 minutes). [Table 5A] [Table 5B] [Table 5C]

[0096] Chymotrypsin enzyme activity measurement using Ala-Ala-Phe-7-amido-4-methylcoumarin

[0097] A cold HCl solution of approximately 2 units of chymotrypsin, a 1.18 mM substrate solution, 2 M calcium chloride, and an 80 mM Tris HCl buffer solution were prepared. The substrate solution and calcium chloride solution were added to 3 mL of the buffer solution, and the mixture was stirred by inversion at approximately 25°C to prepare a blank reaction mixture and a test reaction mixture (Table 6). Furthermore, the HCl solution was added to the blank reaction mixture, and the enzyme solution was added to the test reaction mixture, and the mixture was immediately stirred by inversion. 256 The increasing absorbance at 1000 kJ / min was recorded for 3-5 min. The maximum linear rate of absorbance at least four times at intervals of more than 1 min was used to calculate the A of both the blank and test reaction mixtures. 256 / min. [Table 6]

number

[0098] Measurement of chymotrypsin inactivation in the presence of inhibitors: Chymotrypsin at 1 mM / L was incubated with specific known inhibitors (listed in Table 1 above) and with combinations of metal salts and reducing agents (listed in Table 2 above). Inhibitor inactivation served as a positive control.

[0099] Measurement of oxidative inactivation (activity and pH) of chymotrypsin in the presence of combinations of metal ions and reducing agents.

[0100] For a 200 μl reaction mixture, approximately 10 μl of chymotrypsin was added to a 96-well microtiter plate at 37°C with a combination of metal salts and reducing agents (listed in Table 2, serial numbers 1 to 31) at the respective concentrations in buffer ( aboveTable 3 provides details regarding the use of specific combinations from those provided in Table 2, serial numbers 1 through 31. The incubations were continued for 5, 15, and 30 minutes, followed by the addition of 90 μl of substrate (listed in Table 1A). Enzyme activity was measured spectrophotometrically in a microplate reader and compared to the original activity assay. pH was measured with a Hanna Combination pH electrode (reactions with the enzyme were performed in triplicate). Table 7 below shows the enzymatic activity of chymotrypsin after specific time periods following treatment with the combination of metal salt / complex and reducing agent under evaluation. [Table 7]

[0101] Tables 8A-8C below show the pH values ​​measured for chymotrypsin at different time intervals (i.e., 5 minutes, 15 minutes, and 30 minutes). [Table 8A] [Table 8B] [Table 8C]

[0102] Enzyme activity of carboxypeptidase B using hippurylarginine.

[0103] A cold deionized solution of approximately 4 units of carboxypeptidase B and a solution of 1 mM hippurylarginine in 25 nM Tris HCl buffer (pH 7.65) containing 100 nM sodium chloride were each prepared at approximately 25° C. According to Table 9, 3 mL reaction mixtures (test and blank mixtures) were prepared at 25° C. using the solutions prepared above. Calculation formula:

number

[0104] Measurement of carboxypeptidase B inactivation in the presence of inhibitors: Carboxypeptidase B (1 mM / L) was incubated with specific known inhibitors (listed in Table 1A above) and metal salt / reducing agent combinations (listed in Table 2 above). Inhibitor inactivation served as a positive control.

[0105] Measurement of oxidative inactivation (activity and pH) of carboxypeptidase B in the presence of combinations of metal ions and reducing agents.

[0106] For a 200 μl reaction mixture, approximately 10 μl of carboxypeptidase was added to a 96-well plate. The metal salts and reducing agent combinations (listed in Table 2, serial numbers 1 to 31) were incubated at 37°C in a microtiter plate at the respective concentrations in buffer ( above Table 3 provides details regarding the use of specific combinations from those provided in Table 2, serial numbers 1 through 31. ) Incubation was performed for 5, 15, and 30 minutes, followed by the addition of 90 μl of substrate (listed in Table 1A). Enzyme activity was measured spectrophotometrically in a microplate reader and compared to the original activity assay. pH was measured with a Hanna Combination pH electrode (reactions with enzymes were performed in triplicate). Table 10 below shows the enzymatic activity of carboxypeptidases at specific time periods after treatment with the metal salt / complex and reducing agent combinations under evaluation. [Table 10] TIFF0007744741000021.tif91154

[0107] Tables 11A-11C below show the pH values ​​measured for carboxypeptidase at different time intervals (i.e., 5 minutes, 15 minutes, and 30 minutes). [Table 11A] [Table 11B] [Table 11C]

[0108] Enzyme activity of aminopeptidase M using L-leucine-P-nitroanilide

[0109] A 1 mM tricine solution (prepared in 100 mL of deionized water, Reagent A) and a 50 mM solution of L-leucine-p-nitroanilide in pure methanol (Reagent B) were prepared. Approximately 10 mM L-leucine-p-nitroanilide solution (Reagent C) was prepared by adding 0.1 mL of Reagent B to 4.9 mL of Reagent A. A 200 mM tricine buffer (Reagent D) and a 200 mM tricine buffer containing 0.05% (w / v) BSA, pH 8.0 at 25°C (Reagent E) were prepared in deionized water. A 0.04 Units / mL aminopeptidase solution (enzyme solution, Reagent F) in Reagent E was prepared. Reagent C (Leu-NA, 2.0 ml), Reagent D (200 mM Tricine buffer, 1.0 ml), and deionized water (7.0 ml) were pipetted and mixed in a container by swirling to obtain a reaction cocktail (Reagent G). Reagents G, E, and F were immediately inverted and mixed to prepare test solutions and blank solutions as shown in Table 12. After mixing by stirring, the ΔA 405 nm was recorded and the maximum linear rate was used to obtain A405 / min for the blank and test solutions. Calculation formula:

number

[0110] Measurement of aminopeptidase inactivation in the presence of inhibitors: 1 mM / L of aminopeptidase was incubated with specific known inhibitors (listed in Table 1 above) and metal salt / reducing agent combinations (listed in Table 2 above). Inhibitor inactivation served as a positive control.

[0111] Measurement of oxidative inactivation (activity and pH) of aminopeptidases in the presence of combinations of metal ions and reducing agents.

[0112] For a 200 μl reaction mixture, approximately 10 μl of aminopeptidase was mixed in a buffer solution ( ) with a combination of metal salts and reducing agents (listed in serial numbers 1 to 31 in Table 2) at the respective concentrations at 37°C in a 96-well microtiter plate. above Table 3 provides details regarding the use of specific combinations from those provided in Table 2, serial numbers 1 through 31. The mixtures were incubated for 5, 15, and 30 minutes, after which 90 μl of substrate (listed in Table 1A) was added. Enzyme activity was measured spectrophotometrically in a microplate reader and compared to the original activity assay. pH was measured with a Hanna Combination pH electrode (reactions with the enzyme were performed in triplicate). Table 13 below shows the enzyme activity of aminopeptidases after specific time periods following treatment with the combination of metal salt / complex and reducing agent under evaluation. [Table 13] TIFF0007744741000028.tif90154

[0113] Tables 14A-14C below show the pH values ​​measured for aminopeptidase at different time intervals (i.e., 5 minutes, 15 minutes, and 30 minutes). [Table 14A] [Table 14B] [Table 14C]

[0114] Based on the experiments performed and those described above, it could be concluded that the highest level of inhibition of enzyme activity (proteolysis) was observed by utilizing the following reducing agent and metal salt combinations: sodium ascorbate-vanadium oxide, benzohydroxamic acid-vanadium sulfate, mannitol-vanadium sulfate, uric acid-manganese gluconate, reduced glutathione-chromium chloride, and reduced glutathione-vanadium oxide.

[0115] Bioavailability test

[0116] To protect against protease degradation in the acidic pH environment of the stomach, a capsule-in-capsule formulation was prepared in which the enteric-coated capsule can protect the peptide from the stomach environment, the MIRA granules present on the outside can inactivate proteolytic enzymes, and the permeation enhancer can promote / increase the absorption of the peptide through the epithelial membrane via intestinal permeation.

[0117] Preparation of capsule-in-capsule dosage form

[0118] Preparation of granules containing reducing agents and metal salts

[0119] Tables 15A-15F below show granule formulations (referred to herein as MIRA granules) containing different reducing agents and metal salts that were prepared for bioavailability testing. [Table 15A] [Table 15B] [Table 15C] [Table 15D] [Table 15E] [Table 15F]

[0120] Preparation method of MIRA granules

[0121] The following is the preparation process of MIRA granules.

[0122] A) Preparation of Binder Solution: A 0.3% w / v HPMC E-5 solution was prepared by dissolving 75.0 mg of HPMC E-5 in 25.0 mL of deionized (DI) water. B) Preparation of Powder Mix: All ingredients were placed in a suitable container and thoroughly mixed using a plastic bag to ensure uniformity. C) Preparation of Granules: 2.0 mL of binder solution was added dropwise and manually granulated to prepare granules (2 mL of binder was required to granulate 2.5 g of powder mix). D) Drying of Granules: The granules were placed in a hot air oven and dried at 40°C for 12 hours. (E) Sieving of Granules: The dried granules were sieved using a 40# stainless steel mesh. The sieved granules were collected in a suitable glass container and stored at room temperature. (Note that the temperature and humidity were maintained at 23°C and 39% RH throughout the entire granulation process.)

[0123] Preparation of peptide-containing granules

[0124] Tables 16A-16I below show different peptide-containing granule formulations (referred to herein as PA granules) that were prepared to test bioavailability. [Table 16A] [Table 16B] [Table 16C] [Table 16D] [Table 16E] [Table 16F] [Table 16G] [Table 16H] [Table 16I]

[0125] Method for granulating peptide granules

[0126] A. Preparation of Binder Solution: A 0.3% w / v HPMC E-5 solution was prepared by dissolving 75.0 mg of HPMC E-5 in 25 mL of deionized (DI) water. B. Preparation of Powder Mixture: All ingredients except the liquid excipients were accurately weighed and mixed for 5 minutes using a plastic bag. C. Addition of Binder: The weighed liquid excipients and peptide were added to the 0.3% w / v HPMC E-5 binder solution. The resulting mixture was added dropwise for wet granulation. D. Drying of Granules: The granules prepared by granulation were placed on a silica gel bed in a vacuum desiccator and dried overnight. E. Sieving of Granules: The dried granules were sieved using a 40# stainless steel mesh. The sieved granules were collected in an appropriate glass container and stored at room temperature. (During the entire granulation process, the temperature was maintained at 22°C and the humidity at 35% RH.)

[0127] Capsule filling

[0128] The MIRA and peptide granules were manually filled into capsules using a weighing balance. Tables 17A and 17B below show the capsule sizes used for capsule filling for the rat and dog studies, respectively. [Table 17A] [Table 17B]

[0129] Capsule packaging and storage

[0130] After packaging the capsules in a poly bag, the packaged capsules were transferred into an HDPE (high density polyethylene) container equipped with a silica bag for humidity control.

[0131] Preparation of placebo granules for batch testing

[0132] Placebo batches were prepared using Sunset Yellow and Blue to visually understand the disintegration and release of granules from the capsules. The dried granules were filled into capsule size 3, and the disintegration time was measured using a guide disk in a disintegration tester (Electrolab). The disintegration time was measured to be 3±1 minutes in water and phosphate buffer (pH 6.8) at 37±0.2°C.

[0133] Dissolution test of placebo granules (capsule-in-capsule)

[0134] Outer capsule: Capsule size 0, yellow granules.

[0135] Inner capsule: Capsule size 4, blue granules.

[0136] Dissolution tests were carried out at 37±0.5°C in 900 mL of 0.1 N HCl solution (2 hours) and in pH 6.8 phosphate buffer using a dissolution tester (Electrolab Mumbai).

[0137] The outer enteric-coated capsule remained intact in 0.1 N HCl solution (i.e., was stable in the gastric matrix), but visually observed that it began to disintegrate after 3 minutes in pH 6.8 phosphate buffer.

[0138] The inner capsules began to disintegrate in 8 minutes and were completely dissolved within 13 minutes. Similarly, MIRA granules began to dissolve in alkaline phosphate buffer solution in 3 minutes and the peptide was completely dissolved within 13 minutes.

[0139] Capsule evaluation

[0140] Rat test determination of insulin glargine, octreotide acetate, and teriparatide - 50 mg / 30 mg granules (taken from one capsule) were accurately weighed and dissolved in the mobile phase. The dispersion was sonicated for 5 minutes, filtered using a 0.22 μm syringe filter, and injected into the HPLC. Calibration curves for all APIs were plotted using multiple dilutions in each mobile phase as recommended in USP 2017. The assay was performed, and the drug content was calculated using the calibration curves. [Table 18]

[0141] Stability study of insulin glargine capsules [Table 19]

[0142] Capsule filling - mode: manual filling; capsule size: 2; granule filling weight: same as each formulation shown above.

[0143] Storage of capsules and granules: Temperature: 25±3°C. Humidity: 35±5%RH. Container: HDPE 60 cc for capsules / transparent glass vial with rubber stopper for granules.

[0144] Analytical results: From the formula (y = 30.977x - 292.26 obtained from HPLC data), it was found that the content of insulin glargine present in the formulation was 105.2% and 115.8% for batch I and batch II, respectively.

[0145] 75-day stability study: Both batches were stored at 25°C for 85 days and then re-analyzed.

[0146] Analytical results: From the formula (y = 30.977x - 292.26 obtained from HPLC data), the content of insulin glargine present in the formulation was found to be 97.25% and 91.63% for batch I and batch II respectively, as can be seen from Table 20 below. [Table 20]

[0147] Teriparatide stability study [Table 21]

[0148] Capsule filling - mode: manual filling; capsule size: 2; granule filling weight: same as each formulation shown above.

[0149] Storage of capsules and granules: Temperature: 25±3°C. Humidity: 35±5%RH. Container: HDPE 60 cc for capsules / Transparent glass vial with rubber stopper for granules

[0150] Analytical Results: From the formula (y = 18.924x - 22.539 obtained from HPLC data), the content of teriparatide acetate present in the formulation was found to be 98.02% and 106% for Batch I and Batch II, respectively.

[0151] 75-day stability study: Two batches were stored at 25°C for 75 days and then re-analyzed.

[0152] From the formula (y = 18.924x -22.539 obtained from HPLC data), the content of teriparatide acetate present in the formulation was found to be 92.88% and 89.76% for Batch I and Batch II, respectively. [Table 22]

[0153] Leuprorelin acetate assay in rats

[0154] Preparation of leuprorelin acetate HPLC calibration curve: Approximately 112 mg and 104 mg of granules (equivalent to 100 μg) were accurately weighed and diluted with 1 mL of mobile phase; the theoretical concentration of leuprorelin acetate in this dispersion was found to be 100 μg / mL. The dispersion was vortexed for 2 minutes and then filtered with a 0.2 μm syringe. 20.0 μL of the filtrate was injected into the HPLC system to measure the peptide content. Analysis results: Using the formula y = 42.67x - 76.447 obtained from the HPLC analysis for peptides, the leuprorelin acetate content (% peptide amount) was: (A) leuprorelin acetate + Labrasol ALF Granules (PA2): 97.45%; and (B) leuprorelin acetate + Labrasol ALF + Piperine Granules (PA2+3): 105.11%.

[0155] Dissolution test of leuprorelin acetate capsules

[0156] Based on the analytical results, dissolution tests were conducted on Labrasol ALF and the granule (PA2) formulation containing leuprorelin acetate.

[0157] Setting 1: Use of dialysis membrane - Substrate: phosphate buffer solution at pH 6.80; Volume: 10 mL; Discharge volume: 400 μl; Stirring speed: 100 RPM; Filling granules: 140 mg (equivalent to one capsule); Dialysis membrane specification: HIMEDIA LM395-30MT; Pore size: 25 nm; Average planar width: 29.31 mm; Average diameter: 17.5 mm. [Table 23]

[0158] Setup 2: Using a rotating basket (USP TYPE 1) - Substrate: Phosphate buffer, pH 6.80; Volume: 25 mL; Discharge volume: 400 μl; Stirring speed: 100 RPM; Fill granules: 140 mg in a gelatin capsule. [Table 24]

[0159] Configuration 3: Using a rotating basket (USP TYPE 1)

[0160] Substrate: phosphate buffer at pH 6.80; volume: 30 mL; discharge volume: 400 μl; stirring speed: 500 RPM; filling granules: 140 mg in gelatin capsules. [Table 25]

[0161] Configuration 4: Using a rotating basket (USP TYPE 1)

[0162] Substrate: phosphate buffer solution at pH 6.80; volume: 30 mL; discharge volume: 400 μl; stirring speed: 500 RPM; filling granules: hypromellose capsules (size 0) containing 140 mg. [Table 26]

[0163] Configuration 5: Using a rotating basket (USP TYPE 1)

[0164] Substrate: phosphate buffer solution at pH 6.80; volume: 30 mL; discharge volume: 500 μl; stirring speed: 100 RPM; filled granules: 140 mg of granules were placed directly into the rotating basket without being prepared into capsules. [Table 27]

[0165] Setting 6: Using a magnetic stirrer

[0166] Substrate: phosphate buffer solution at pH 6.80; volume: 30 mL; discharge volume: 500 μl; stirring speed: 200 RPM; filling granules: prepare 140 mg in gelatin capsules (capsule size 2). [Table 28]

[0167] Setting 7: Using a magnetic stirrer

[0168] Substrate: phosphate buffer solution at pH 6.80; volume: 30 mL; discharge volume: 500 μl; stirring speed: 200 RPM; filling granules: prepare 140 mg in gelatin capsules (capsule size 2). [Table 29]

[0169] From the data in Tables 24-29, it can be noted that leuprorelin acetate did not dissolve from the granules and pass through the dialysis membrane (Setting 1). In Setting 2, a rotating basket (USP TYPE 1) was used, and the basket rotation was unable to generate enough rotational force to move / rotate the material in the matrix, causing the material to settle with the gelatin, ultimately affecting dissolution. In Setting 3, the basket RPM was increased (from 100 to 500), but the CDR% remained at 24.75% even with the increased basket rpm. In setting 4, the rotating basket RPM was increased and hypromellose capsules were used instead of gelatin capsules, resulting in a CDR% of 29.97%; in setting 5, the rotating basket was used without capsules, resulting in a CDR% of 90%, demonstrating that the materials (gelatin / hypromellose) can increase viscosity and delay the release of leuprorelin acetate from the granules; in settings 6 and 7, a magnetic stirrer was used to ensure good rotation of the substrate throughout the analysis, resulting in CDR% of 108.8% and 102.6%, respectively; all experiments were performed at 37°C, but changes in temperature can affect capsule disintegration. The disintegration time at 25°C was 12 minutes, and at 37°C it was less than 2 minutes.

[0170] Determination of liraglutide sodium using HPLC

[0171] Test 1: Procedure: Approximately 10.0 mg of Granules PA2 (Table 16F) and Granules PA3+4 (Table 16G) were diluted in 10.0 mL of HPLC diluent (10% ACN in DI water). The theoretical concentration of liraglutide sodium in this dispersion was found to be 74 μg / mL. The dispersion was sonicated in an ultrasonic bath for 5.0 minutes, then filtered through a 0.2 μm syringe. 20.0 μL of the filtrate was injected into the HPLC system to measure the peptide content. Analysis Results: From the formula y = 79.283x - 571.72 obtained from the HPLC for peptide, the liraglutide sodium formulation with Labrasol ALF (PA2) had a content of 44.4%, and the liraglutide sodium formulation with piperine and Solutol HS15 (PA3+4) had a content of 25.4%. (a) After vortexing for 2 minutes, the assay result of the liraglutide sodium formulation with piperine and Solutol HS15 was 67.39%; (b) After vortexing for 2 minutes, the assay result of the liraglutide sodium granule formulation with Labrasol ALF was 80.43%; (c) After vortexing for 5 minutes, the assay result of the liraglutide sodium formulation with piperine and Solutol HS15 was 95.66%; (d) After vortexing for 5 minutes, the assay result of the liraglutide sodium granules with Labrasol ALF was 96.99%.

[0172] Liraglutide dissolution test

[0173] Substrate: phosphate buffer solution at pH 6.80; volume: 30 mL; discharge volume: 500 μl; stirring speed: 200 RPM; [Table 30] [Table 31]

[0174] Analysis results: The CDR% of liraglutide granules PA and PA3+4 were 112.85% and 93.64%, respectively.More than 50% of liraglutide sodium was dissolved between 5 and 7 minutes.

[0175] Quantitative analysis of glucose and insulin glargine contents in plasma of STZ-induced diabetic rats

[0176] After administering insulin glargine to the mid-jejunum of STZ-induced diabetic rats, the glucose and insulin glargine contents in their plasma were quantified.

[0177] Test Formulation I: Insulin glargine (Lantus (R) )-Appearance: Injection solution in a prefilled pen syringe; Concentration: 100 IU / ml; Storage conditions: 2-8°C; Dose: 0.2 U / kg; Route of administration: SC.

[0178] Test Formulation II: Insulin glargine formulation - Granules MIRA1 (reduced glutathione / chromium picolinate from Table 15A) + permeation enhancer 1 (PA1 from Table 1) (oral solution in TRIS buffer) - Dose: 1.7 U / animal; Route of administration: mid-jejunum.

[0179] Test Formulation III: Insulin glargine formulations - PA2 (as per Table 16B) and MIRA2 (sodium ascorbate / vanadium oxide as per Table 15B) (oral solution in buffer) - Dose: 1.7 U / animal; Route of administration: mid-jejunum.

[0180] Test results: No rats died in either the subcutaneous injection or mid-jejunal administration. Clinical symptoms were normal. Regarding sampling time points, blood was collected at 0, 20, 40, 60, 120, and 150 minutes after administration during administration. Approximately 100 μl of blood was collected by retro-orbital sinus puncture into a pre-filled Na-EDTA Eppendorf flask. The blood was centrifuged at 5000 rpm for 5 minutes at 4°C to obtain plasma. Blood glucose was measured using a blood glucose meter immediately after collection. [Table 32] [Table 33]

[0181] ELISA test

[0182] Test materials: Insulin glargine ELISA kit (Invitron Ltd, Cat. No. MBS495369).

[0183] Principle: This glargine ELISA is a two-site immunoassay that uses a monoclonal antibody immobilized on the wells of a microtiter plate and a soluble antibody conjugated with horseradish peroxidase (HRP). Plasma samples were incubated together in the wells of the microtiter plate and washed, followed by the addition of an HRP-conjugated antibody solution. Before measurement, unbound HRP-conjugated antibody was washed away and then a secondary incubation was performed. An enzyme substrate was added to each well of the microtiter plate, and after a short incubation, the reaction was stopped by the addition of additional reagents. The intensity of the color developed in each well was quantified using a microtiter plate reader set to record transmitted light at a wavelength of 450 nm (using the kit protocol recommended by the manufacturer, catalog number MBS495369).

[0184] Procedure: Allow all kit components and samples to stand at room temperature before use. Secure the required number of coated strips to the plate holder. Store any strips not being used immediately in a sealed polyethylene bag containing silica gel desiccant. Ensure that any remaining space on the plate holder is filled with uncoated strips to ensure uniform heat transfer during incubation. Pipet 100 μl of sample buffer into each well. Pipet 25 μl of standard or sample into each well. It is recommended that each standard and sample be tested in duplicate. Cover the plate holder and incubate for 2 hours at room temperature (18-22°C). Remove the plate holder cover and use an automated plate holder washer to wash three times using a combination of chilled working strength wash buffer (300 μl per cycle). Pipet 100 μl of working strength conjugated antibody into each well. Cover with a plate sealer and incubate for 4 hours at 4°C (2-8°C). Remove the plate sealer and wash three times with chilled* working strength wash buffer using an automated plate holder washer. Next, add 100 μl of substrate solution to each well and incubate for 15 minutes in the dark at room temperature (18-22°C). Add 100 μl of stop solution to each well. Measure light transmittance using a microtiter plate reader set to 450 nm, with background subtraction measured at OD 620 / 650 nm, if possible. Figure 1 shows a graph depicting the concentration versus time profile of insulin glargine (mU / L) from different formulations.

[0185] The test formulation insulin glargine-MIRA1 (reduced glutathione / chromium picolinate) plus permeation enhancer 1 (PA1) was found to have a relative bioavailability of 9.25%, and the test formulations insulin glargine-PA2 and MIRA2 (sodium ascorbate / vanadium oxide)) were found to have a relative bioavailability of 28.86%.

[0186] Quantitative analysis of leuprorelin acetate content in dog plasma using an ELISA kit

[0187] Standard test formulation: Leuprorelin acetate: LUPRODEX; Depot) - Concentration: Each vial contains 3.75 mg of leuprorelin acetate; Date of manufacture: November 2017; Expiration date: October 2020; Storage environment: Store at room temperature (below 25°C), do not freeze; Number of vials: 1 vial with diluent.

[0188] Test formulation FB: MIRA5 (Table 15E) + PA2 (Table 16H) - Appearance: hard gelatin capsule with a white capsule cap and a white capsule body; Concentration: each capsule contains 300 mg and 1.25 mg of leuprorelin acetate; Date of manufacture: April 21, 2018; Expiration date: not provided; Storage environment: stored at room temperature (below 25°C); Number of test capsules: 70 capsules per container.

[0189] Test Formulation H: MIRA2 (Table 15F) + PA2 + 3 (Table 16I) - Appearance: Hard gelatin capsule with a white cap and a white capsule body; Concentration: Each capsule contains 300 mg and 1.25 mg of leuprorelin acetate; Date of manufacture: April 19, 2018; Expiration date: None; Storage environment: Store at room temperature (below 25°C); Number of test capsules: 70 capsules per container. [Table 34]

[0190] During the dosing period, dogs (Canidae; breed; Beagle) were fasted overnight from 12 hours before dosing through 4 hours after dosing (water was allowed). 720 hours after dosing, the dogs were observed for any adverse events. Each dog's body weight was measured and recorded before dosing.

[0191] Sample sampling time

[0192] Approximately 2 mL of blood samples were collected from the jugular vein of each subcutaneously and orally dosed dog at 0, 1, 2, 6, 12, 24, 48, 72, 96, 120, 240, 312, 360, 480, and 720 hours (a total of 15 sampling time points) into K2EDTA-coated and labeled sampling tubes.

[0193] ELISA test conditions

[0194] Test material: Catalog number S-1174 (Des-Gly10, D-Leu6, Pro-NHEt9)-LHRH (leuprolide). Kit protocol: Manufacturer's recommended protocol used (Cat. number S1174).

[0195] Test results: The relative bioavailability of test formulation FB was 56.53%, and the relative bioavailability of test formulation H was 16%.

[0196] Determination of liraglutide content in dog plasma using an ELISA kit

[0197] Test Formulation I: Liraglutide - Appearance: Injection solution prefilled in a prefilled pen; Concentration: 6 mg / ml; Date of manufacture: February 2017; Expiration date: July 2019; Storage environment: 2-8°C; Dose: 0.6 mg per dog; Route of administration: SC

[0198] Test Formulation II (FA): MIRA5 (Table 15E) + PA2 (Table 16F) - Dose: 12 mg (1 capsule) / dog; Route of administration: oral.

[0199] Test Formulation III(G): MIRA5 (Table 15E) + PA3 + 4 (Table 16G) - Dose: 12 mg (1 capsule) / dog; Route of administration: oral. [Table 35]

[0200] No dog deaths occurred after subcutaneous or oral liraglutide administration. Clinical symptoms were normal. The weight of each dog was measured and recorded before the study. During the dosing period, dogs (Canidae; breed; Beagle) were fasted overnight from 12 hours before dosing until 4 hours after dosing (water was allowed). During the dosing period, blood samples were collected from the dogs at time points of 0, 20, 30, 60, 120, 180, 240, and 480 minutes after dosing. 2 mL of blood samples were collected from the jugular vein into K2EDTA-coated, labeled sample collection tubes. Glucose was measured using a blood glucose meter immediately after blood sampling.

[0201] ELISA test conditions

[0202] Test materials: ELISA kit (Krishgen BioSystems, Cat. No. KBI5020 Ver. 2.0).

[0203] Kit protocol: The manufacturer's recommended protocol was used (Cat. No. KBI5020 Ver. 2.0). (1) Define the wells for the diluted standard, blank control, and sample, preparing five wells for the standard and one well for the blank control. Add 50 μL of the standard (reading reagent preparation), blank control, and sample to the appropriate wells. Immediately add 50 μL of liraglutide-biotin to each of the above wells. Gently shake the plate (use of a plate shaker is recommended). Cover with a plate cover. Incubate at 37°C for 1 hour. Liraglutide-biotin may appear cloudy. Leave at room temperature and mix gently until the solution is homogenous. (2) Aspirate the solution from the microplate, then wash each well with 350 μL of 1x washing solution using a spray bottle, multichannel pipette, manifold dispenser, or automatic washer. Allow to stand for 1-2 minutes. Invert the plate onto absorbent paper and press to remove any remaining liquid. Repeat three times. After the final wash, aspirate or pour off any remaining wash solution, then invert the microplate onto absorbent paper. (3) Add 100 μL of streptavidin-HRP sample to each well. Cover with a plate sealer and incubate at 37°C for 30 minutes. (4) Repeat the aspirate / wash procedure as performed in step (2) a total of five times. (5) Add 90 μL of substrate solution to each well. Cover with a new plate sealer. Incubate at 37°C for 10-20 minutes (not to exceed 30 minutes). Protect from light. The solution will turn blue upon addition of the substrate solution. (6) Add 50 μL of stop solution to each well. The solution will turn yellow upon addition of the stop solution. Tap the side of the plate to mix the mixture. If the color change of the solution is uneven, gently tap the microplate to ensure the solution is thoroughly mixed. (7) Remove any water droplets or fingerprints from the bottom of the microplate and ensure that there are no air bubbles on the liquid surface in the wells.Measurements are taken at 450 nm using a microplate reader.

[0204] The relative bioavailability of test formulation FA (Mira5 and liraglutide + Labrasol) was found to be 3.82%, and the relative bioavailability of test formulation G (Mira5 and liraglutide + Solutol HS15 + piperine) was found to be 3.57%.

[0205] Quantitative analysis of octreotide content in rat plasma by ELISA

[0206] Test Formulation I: Octreotide - Appearance: Injection solution; Concentration: 0.1 mg / ml; Storage condition: 2-8°C; Dose: 10 μg / kg; Administration route: SC

[0207] Test Formulation II: MIRA3 (Table 15C) + PA1 (Table 16C) was administered to the distal small intestine (ileum) of anesthetized SD rats; dose: 144 μg / animal; route of administration: distal small intestine (ileum) injection. The rats were not fasting during the test. [Table 36]

[0208] No rats died after subcutaneous or distal small intestinal (ileal) administration of octreotide. Clinical symptoms were normal. During the administration period, blood samples were collected from dogs at 0, 7, 15, 30, 45, 60, and 90 minutes after administration. Blood samples were collected by retro-orbital venous plexus puncture, and approximately 100 μl of blood was collected in an Eppendorf tube prefilled with Na-EDTA. Plasma was obtained by centrifuging the blood samples at 5000 rpm for 5 minutes at 4°C.

[0209] ELISA test

[0210] Test equipment: ELISA kit (Peninsula Laboratories International, Inc., Cat. No. S-1341.0001).

[0211] Kit protocol: The manufacturer's recommended protocol was used (Cat. No. S-1341.0001). Add 25 μl of antiserum (in EIA buffer) to each well of the immunoplate. Add 25 μl of EIA buffer to blank wells and incubate at room temperature for 1 hour. Add 50 μl of each standard or sample (in diluent). Do not wash the plate before adding. Add 50 μl of diluent to blank wells and incubate at room temperature for 2 hours. A short preincubation may result in reduced sensitivity. Rehydrate Bt-tracer (in EIA buffer) and add 25 μl to each well and incubate at 4°C overnight. For best results, return to RT before proceeding. Wash the immunoplate five times with 300 μl / well of EIA buffer. Take care to avoid cross-contamination between wells during the first wash / dispensing. During each wash cycle, empty the contents of the plate with a quick, flick of the wrist, then gently blot the top of the plate dry on a paper towel. After dispensing 300 μl of EIA buffer into each well, gently rock the microplate for at least a few seconds. This wash step is essential. Add 100 μl of streptavidin-HRP to each well. Tap or centrifuge the SAHRP tube to collect all liquid components at the bottom, dilute the liquid inside 1 / 200 (60 μl / 12 mL) in EIA buffer, and vortex. Add 100 μl to all wells, including the blank. Incubate at room temperature for 1 hour. Wash the immunoplate five times (see step (7)). Add 100 μl of TMB solution per well. Add to all wells, including the blank. Incubate at room temperature (typically 30-60 minutes). Read the blue color development at 650 nm and perform calculations using the data. The reaction is stopped by adding 100 μl of 2N HCl to each well. The absorbance is read at 450 nm within 10 minutes.

[0212] The test formulation MIRA3 (uric acid:sodium vanadate) + PA1 had a bioavailability of 0.41%.

[0213] Determination of teriparatide content in rat plasma by ELISA

[0214] Test Formulation I: Teriparatide; Appearance: Injectable solution; Concentration: 600 μg / 2.4 ml; Storage condition: 2-8°C; Dose: 10 μg / animal; Administration route: SC

[0215] Test Formulation II: MIRA4 (Table 15D) and PA1 (Table 16D) were administered to the distal small intestine (ileum) of anesthetized rats at a dose of 240 μg / animal; Dose: 240 μg / animal; Route of administration: distal small intestine (ileum).

[0216] Test Formulation III: MIRA1 (Table 15A) and PA3 (Table 16E) were administered to the distal small intestine (ileum) of anesthetized SD rats at a dose of 240 μg / animal; dose: 240 μg / animal; route of administration: distal small intestine (ileum). [Table 37]

[0217] The animals were not fasted during the experimental period. No rats died after subcutaneous or distal small intestinal (ileal) administration of teriparatide. Clinical symptoms were normal. During the administration period, blood samples were collected from dogs at 0, 7, 15, 30, 45, 60, and 90 minutes after administration. Blood samples were collected by retroorbital venous plexus puncture, and approximately 100 μl of blood was collected in an Eppendorf tube prefilled with Na-EDTA. Plasma was obtained by centrifuging the blood samples at 5000 rpm for 5 minutes at 4°C.

[0218] ELISA test

[0219] Test material: ELISA kit (Immutopics Cat. No. 60-3900)

[0220] Kit protocol: The manufacturer's recommended protocol was used (Cat. No. 60-3900). Place a sufficient number of streptavidin-coated strips into the holder and test for parathyroid hormone (PTH) standards, controls, and unknown samples; pipet 150 μl of each standard, control, and sample into each designated or mapped well. Freeze the standard and control samples immediately after use. Pipet 50 μl of working antibody diluent (1 part HRP antibody and 1 part biotinylated antibody) into each well; cover with a plate sealer and aluminum foil to protect from light; incubate the plate for 3 hours at room temperature in a horizontal rotor set at 180-200 rpm. Remove the aluminum foil and plate sealer, and aspirate the contents of each well using an automated microplate washer. Wash each well five times with 350 μL of working wash diluent, completely aspirating the contents of each well. An appropriate vacuum device can be used; pipet 200 μl of ELISA HRP substrate into each well; re-cover the plate with a plate sealer and aluminum foil. Incubate for 3 hours at room temperature on a horizontal rotor set at 180-200 rpm; remove the aluminum foil and plate sealer. Within 5 minutes, read the absorbance at 620 nm (see inscription) on a microplate reader using the 0 pg / mL standard well as a blank; immediately pipet 50 μL of ELISA stop solution into each well. Mix for 1 minute on a horizontal rotor; within 10 minutes, read the absorbance at 450 nm using a microplate reader using 200 μL of substrate and 50 μL of stop solution as a reagent blank; if dual wavelength compensation is possible, set the measurement wavelength to 450 nm and the reference wavelength to the absorbance used in step #9.

[0221] The relative bioavailability of the test formulations MIRA4 (sodium ascorbate:manganese gluconate) and PA1 was 0.89%. The bioavailability of the test formulations MIRA1 (reduced glutathione / chromium picolinate) and PA3 was 0.89%.

[0222] While various embodiments of the present invention have been described and illustrated above, other and further embodiments of the present invention may be made without departing from the basic scope thereof. The scope of the present invention is determined by the claims. The present invention is not limited to the described embodiments, variations, or examples, but also includes those that, when combined with information and knowledge available to those skilled in the art, enable one skilled in the art to make and use the invention.

[0223] The present disclosure provides pharmaceutical compositions that can overcome the drawbacks associated with compositions reported in the background art.

[0224] The present disclosure provides pharmaceutical compositions for effective peptide delivery.

[0225] The present disclosure provides pharmaceutical compositions for oral peptide delivery.

[0226] The present disclosure provides pharmaceutical compositions that, upon oral ingestion, at least partially protect peptides from proteolytic degradation.

[0227] The present disclosure provides pharmaceutical compositions that increase the bioavailability of peptides.

[0228] The present disclosure provides a pharmaceutical composition with safety.

[0229] The present disclosure provides pharmaceutical compositions that are cost-effective, easy to prepare, and have a long shelf life.

Claims

1. a pharmaceutically effective dose of at least one peptide, and a pharmaceutically acceptable dosage of (a) a metal salt or metal complex, in combination with (b) at least one reducing agent; the (a) metal salt or metal complex is selected from the group consisting of vanadium(V) oxide, sodium vanadate, and vanadium sulfate, and the (b) at least one reducing agent is sodium ascorbate, benzohydroxamic acid, mannitol, cysteine, or reduced glutathione; and the combination of the (a) metal salt or metal complex and the (b) at least one reducing agent is used to at least partially protect the at least one peptide from proteolytic degradation upon ingestion; A pharmaceutical composition prepared in the form of either an oral solid or oral liquid dosage form (excluding those containing unesterified lutein, zeaxanthin, vitamin C, vitamin E, zinc, and copper).

2. 10. The pharmaceutical composition of claim 1, wherein the vanadium(V) oxide, sodium vanadate, or vanadium sulfate is present in an amount ranging from about 0.01 mg to about 15 mg per unit dose.

3. 2. The pharmaceutical composition of claim 1, wherein the at least one peptide has a molecular weight of 60 kDa or less.

4. The at least one peptide may be insulin, an insulin analog, insulin lispro, insulin peglispro, insulin aspart, insulin glulisine, insulin glargine, insulin detemir, protamine-containing intermediate-acting (NPH) insulin, insulin degludec, B29K(N(ε)hexadecandioyl-γ-L-Glu) A14E B25H desB30 human insulin, B29K(N(ε)octadecandioyl-γ-L-Glu-OEG-OEG) desB30 human insulin, B29K(N(ε)octadecandioyl-γ-L-Glu) A14E B25H desB30 human insulin, B29K(N(ε)eicosandioyl-γ-L-Glu) A14E B25H desB30 Human insulin, B29K (N(ε)octadecandioyl-γ-L-Glu-OEG-OEG) A14E B25H desB30 hu Human insulin, B29K (N(ε)eicosandioyl-γ-L-Glu-OEG-OEG) A14E B25H desB30 Human insulin, B29K (N(ε)eicosandioyl-γ-L-Glu-OEG-OEG) A14E B16H B25H desB30 Human insulin, B29K (N(ε)hexadecandioyl-γ-L-Glu) A14E B16H B25H desB30 Human insulin, B29K (N(ε)eicosandioyl-γ-L-Glu-OEG-OEG) A14E B16H B25H desB30 Human insulin, B29K (N(ε)octadecandioyl) A14E B25H desB30 human insulin, GLP-1, GLP-1 analogues, acylated GLP-1 analogues, diacylated GLP-1 analogues, semaglutide, liraglutide, exenatide, lixisenatide,Dual agonists of GLP-1 receptor and glucagon receptor, amylin, amylin analogs, pramlintide, somatostatin analogs, octreotide, lanreotide, pasireotide, goserelin, buserelin, leptin, leptin analogs, metreleptin, peptide YY, peptide YY analogs, glatiramer, leuprorelin, teriparatide, desmopressin, human growth hormone, human growth hormone analogs, glycopeptide antibiotics Glycosylated cyclic or polycyclic nonribosomal peptide antibiotics, vancomycin, teicoplanin, telavancin, bleomycin, ramoplanin, decaplanin, bortezomib, cosyntropin, chorionic gonadotropin, menotropin, sermorelin, luteinizing hormone-releasing hormone, somatropin, calcitonin, salmon calcitonin, pentagastrin, oxytocin, nesiritide, anakinra, enfuvirtide, pegvisomant, dorna 2. The pharmaceutical composition of claim 1, wherein the active ingredient is selected from the group comprising pegylated interferon alfa, lepirudin, anidulafungin, eptifibatide, interferon alfacon-1, interferon alfa-2a, interferon alfa-2b, interferon beta-1a, interferon beta-1b, interferon gamma-1b, pegylated interferon alfa-2a, pegylated interferon alfa-2b, pegylated interferon beta-1a, fibrinolysin, vasopressin, aldesleukin, epoetin alfa, darbepoetin alfa, epoetin beta, epoetin delta, epoetin omega, epoetin zeta, filgrastim, interleukin-11, cyclosporine, glucagon, urokinase, viomycin, thyrotropin-releasing hormone, leucine enkephalin, methionine enkephalin, substance P, adrenocorticotropic hormone, parathyroid hormone, or a pharmaceutically acceptable salt thereof.

5. 2. The pharmaceutical composition of claim 1, wherein the at least one peptide and the metal salt or metal complex are physically separated in the pharmaceutical composition.

6. 2. The pharmaceutical composition of claim 1, wherein the at least one peptide and the metal salt or metal complex are present in separate compartments.

7. 10. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is in a capsule-in-capsule dosage form or a tablet-in-capsule dosage form.

8. 10. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition comprises at least one reducing agent in an amount ranging from about 1 mg to about 1000 mg per unit dose.

9. 10. The pharmaceutical composition of claim 1, further comprising at least one absorption enhancer, wherein the absorption enhancer is present in an amount ranging from about 10 mg to about 1000 mg per unit dose.

10. The pharmaceutical composition of claim 1, comprising less than about 5% (v / v) water.

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